1.5 Long hair should be tied back neatly, away from the shoulders.1.6 Enclosed footwear must be worn – (thongs and open sandals are not allowed).1.7 Avoid placing any object in your mouth – (pencils, pens, fingers etc).1.8 Cover any open cuts on hands and other exposed skin surfaces and/or wear gloves.

What is safety hazard in histopathology laboratory?

Sharp tools Changing the scalpel blade and handling these tools can be dangerous if not done correctly. The cutting blade inside of a cryostat or a microtome is extremely sharp and many injuries can occur when working with these instruments or when performing maintenance.

What are the 3 main categories of hazards in a histopathology laboratory?

Different types of hazards – An important first step in protecting worker health and safety is recognizing workplace hazards. Most hazards encountered fall into three main categories: chemical, biological, or physical. Cleaning agents and disinfectants, drugs, anesthetic gases, solvents, paints, and compressed gases are examples of chemical hazards.

  • Potential exposures to chemical hazards can occur both during use and with poor storage.
  • Biological hazards include potential exposures to allergens, infectious zoonotics (animal diseases transmissible to humans), and experimental agents such as viral vectors.
  • Allergens, ubiquitous in animal research facilities, are one of the most important health hazards, yet they are frequently overlooked.

The final category contains the associated with research facilities. The most obvious are slips and falls from working in wet locations and the ergonomic hazards of lifting, pushing, pulling, and repetitive tasks. Other physical hazards often unnoticed are electrical, mechanical, acoustic, or thermal in nature.

What precautions should be taken during tissue processing?

GUIDELINES FOR PROCESSING SPECIMENS WITH KNOWN/PROBABLE INFECTIOUS DISEASE – Specimens from patients with infections not posing a risk to immunocompetent individuals (e.g., routine bacterial and fungal infections, opportunistic pathogens) can be processed as for other pathology specimens using universal precautions.

  1. Specimens from patients with infections (or suspected infections) posing a greater risk to pathology personnel (TB, HBV, HCV, HIV, Creutzfeldt-Jakob disease) must be handled with special precautions.
  2. All specimens must be fixed as soon as possible and stored in rigid leakproof containers.
  3. Gloves must always be worn when handling specimens.

Fresh tissues are potentially infective and all specimens are placed in fixative as soon as possible. Formalin is effective for inactivating viruses (including HIV and HBV) and will reduce the infectivity of mycobacteria. Procedures that could aerosolize an infectious agent (e.g., cutting a specimen with a bone saw) should not be performed.

  • Creutzfeldt-Jakob disease requires special procedures for handling it safely (see specific section).
  • Small specimens (e.g., colon biopsies and open lung biopsies) are usually of immediate diagnostic importance and can be processed as usual as long as the specimens fix in formalin for at least four to six hours.

Larger specimens, if of no immediate diagnostic importance (e.g., a placenta from a normal delivery or a colon resection for trauma) can be sectioned thinly and placed in an adequate volume of fixative (1:10 specimen/formalin fixative ratio) for 72 hours before submitting for histologic processing.

If the specimen is of immediate diagnostic importance, small sections can be cut for blocks and fixed as above before processing. Potentially infectious cases are not photographed in the fresh state. If it is an especially interesting case, pictures after fixation may be taken if special precautions are used in order not to contaminate surfaces or the camera.

Frozen sections on potentially infectious cases may be performed but should be avoided if cytologic preparations can be used or an intraoperative diagnosis is not necessary. Freezing does not inactivate infectious agents. If an infectious case is cut in a cryostat, the cryostat should be decontaminated.

What is the risk in a tissue culture laboratory and safety?

Common hazards are accidental punctures with syringe needles or other contaminated sharps, spills and splashes onto skin and mucous membranes, ingestion through mouth pipetting, and inhalation exposures to infectious aerosols.

What is the most common processing problem in histopathology?

As mentioned throughout this course, proper tissue processing hinges on a variety of factors and variables that must be addressed in order for the outcome to be consistent and free of artifacts. Accurately troubleshooting tissue processing is a mark of a well-seasoned technician, since most troubleshooting skills are gained through first-hand experience.

The most evident processing problem in histology laboratories is under-processed tissue samples. Although some tissues suffer from incomplete fixation, which in turn may lead to improper dehydration, clearing, and infiltration, troubleshooting fixation is best saved for a detailed course on tissue fixation.

Processing problems may be noticed at various points from embedding to slide quality control (QC). Histotechnologists and pathologists have the responsibility of noting any problems seen with the tissue and reporting it to the laboratory supervisor for correction.

  • With each problem there may be several sources to troubleshoot.
  • Starting with the simplest solution first not only takes the least effort, but may save a lot of time.
  • Troubleshoot by process of elimination, from least labor intensive to the most involved solution to the problem.
  • Verifying that the correct processing program was used is a simple solution, compared to changing out all of the processing reagents.

The table below provides some troubleshooting tips for poor processing:

Problem Possible Causes Corrections
Tissue feels soft or mushy during embedding

Tissue may have been grossed in too thickTissue may have been processed on a program that was too short for that tissue type Processing reagents may be saturated with water Paraffin may be saturated with xylene or isopropanol

Reprocess tissue on proper program Reprocess tissue on correct processing protocol Change reagents and reprocess tissue Change paraffin and reprocess tissue

Tissue bounces out of paraffin block during microtomy or tissue does not adhere to block or slides (Commonly experienced with uterus and prostate tissue, as well as dense organ core samples)

Poor dehydration and paraffin infiltration due to water left in the tissue

Change reagents and reprocess tissue on proper processing protocol

Tissue looks greasy and “explodes” or separates rapidly when ribbon is placed on water bath

If the temperature of the water bath is between 45-50° C, then the tissue is under-processed Tissue may have been grossed in too thick Tissue may have been processed on a program that was too short for that tissue type Processing reagents may be saturated with water Paraffin may be saturated with xylene or isopropanol

Reprocess tissue on correct processing protocol Reprocess on proper program Reprocess tissue on correct processing protocol Change reagents and reprocess Change paraffin and reprocess tissue

Tissue does not adhere to slide or falls off easily

If tissue slides are placed in oven prior to deparaffinization in xylene, tissue is under-processed Reagents saturated with water or contaminated with the preceding reagent

Reprocess tissue on correct processing protocolChange reagents and paraffin and reprocess tissue on proper processing protocol

Hematoxylin and eosin (H&E) stained tissue section shows uneven nuclear staining and “blue blobs” lacking distinct chromatin patterns

If tissue was fixed properly, then sample was improperly dehydrated and infiltrated with paraffin

Change reagents and reprocess tissue on proper processing protocol

What happens in a histopathology laboratory?

Why is histopathology important? – Histopathologists are doctors who work closely with other clinical specialties. They can reach a diagnosis by examining a small piece of tissue from the skin, liver, kidney or other organ. This is called a biopsy. They examine the tissue carefully under a microscope, looking for changes in cells that might explain what is causing a patient’s illness.

What is quality control in histopathology laboratory?

Therefore for the histopathology laboratory, quality control must be seen as the policies, practices and procedures enshrined for the production of high quality sections with accurate, timely and complete reports in a cost effective manner at all times.

What are the general hazards in a laboratory?

Hazard Recognition and Solutions – The laboratory environment can be a hazardous place to work. Laboratory workers are exposed to numerous potential hazards including chemical, biological, physical and radioactive hazards, as well as, musculoskeletal stresses.

Why quality control is important in histopathology laboratory?

Histology laboratories must provide high quality services by enforcing a technical quality control system. This can only be achieved by provided accurate, relevant, precise and comprehensive data, which is applied to medical management of patients.

What are the common problems encountered in tissue processing?

Faults in Processing During the processing and cutting of sections, several difficulties may be encountered, due to some faults which may have been made in the previous procedures. A good medical technologist should be alert in taking note of such faults, which, if not immediately corrected shall cause entire failure in the processing, poor sectioning, and ultimately improper evaluation of the tissue in question.

Brittle or hard tissue Prolonged fixation Tissues may be softened by soaking in a small dish or bowl containing water with detergent, phenol or Molliflex.
Prolonged dehydration
Prolonged clearing
Prolonged paraffin infiltration in overheated paraffin oven
Drying out of tissue before actual fixation
Failure to give special handling of tissues when treated with routine processing methods
Clearing agent turns milky as soon as tissue is placed in it Water not completely removed due to incomplete dehydration Repeat dehydration with absolute alcohol, then clear again.
On trimming, tissue smells of clearing agent Clearing agent not completely removed due to insufficient impregnation Block is trimmed down nearest to the tissue, remaining wax is melted on embedding oven and paraffin impregnation is repeated, changing the paraffin at least once before blocking.
Tissue is opaque, section–cutting is difficult due to presence of alcohol Insufficient clearing Repeat clearing; if object has already been embedded, prolong clearing up to 12 hours then re–embed
Tissue shrinks away from wax when trimmed Insufficient dehydration, therefore incomplete clearing and impregnation Repeat the whole procedure
Tissue is soft when block is trimmed Incomplete fixation Repeat fixation
Airholes found on tissue during trimming Incomplete impregnation Repeat impregnation
On trimming, wax appears crystalline Contaminated wax Re–embed in freshly filtered wax
Paraffin block after cooling is moist and crumbles Insufficient paraffin impregnation Repeat paraffin impregnation then re–embed
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FAULTS OBSERVED DURING SECTION – CUTTING Difficulties encountered during cutting of sections are mostly due to faults encountered during the processing of tissues or due to some faults in the technique or cutting itself, and therefore must be evaluated and corrected on a case to case bases, if good tissue sections are to be made.

Sections fail to form ribbons Surfaces and edges of the block are not parallel Re–trim the block
Horizontal surface of the block is not parallel to the knife Re–adjust and re–orient
Paraffin wax is too hard Coat horizontal edges of the block with wax of lower melting point
Knife is tilted too much Reduce the tilt
Sections are too thick Readjust the thickness of the sections
Knife is dull Hone and strop
Sections roll up on cutting so that they adhere and get broken against the knife edge Knife is blunt Sharpen the knife
Tilt of knife is too great Reduce the tilt
Knife edge is dirty Clean the knife edge
Ribbon is curved, crooked or uneven instead of straight Blunt or dull spot on the knife producing an irregular knife edge Adjust the knife so that knife edge will present a uniformly sharp edge to the block; or sharpen
Edges of the block are not parallel but round or wedge shaped Re–trim the block
Knife is not parallel to the block Readjust the knife and the block
Paraffin is impure Repeat impregnation using pure wax
Sections are compressed, wrinkled or jammed Knife is blunt or dull Re–sharpen the knife
Paraffin block is warm and soft Cool the block on ice water until firm
Knife edge is coated with paraffin Clean the knife edge
Sections are too thin Readjust thickness of section
Microtome set screw is loose Tighten the screw
Tilt of knife is too vertical Reduce the tilt
Sections are torn and crumble when cut Incomplete dehydration, clearing, and/or infiltration tissue with wax Remove paraffin with clearing of agent, pass thru decreasing grade of alcohol, then repeat dehydration, clearing and embedding
Paraffin is warm and soft Cool and harden paraffin in ice for ¼ to ½ hour
Knife is blunt Sharpen the knife
Sections are squashed (width of each section less than that of block) Bevel of knife is lost due to incorrect sharpening Re–sharpen, using a knife back or automatic knife sharpener
A hole is formed in the section Bubble or dirt formed in the embedding medium Re–embed in freshly filtered wax if necessary
Hard spot in tissue possibly due to calcium Once embedded in paraffin wax, decalcification is impractical; use base–sledge microtome with a wedge knife
Selections of unequal thickness are produced Tilt of knife is too great or bevel is not cleared, hence object is compressed against the knife edge Reduce the tilt
Clamp set screw on knife or blockholder is loose Tighten screw
Blocks are too large Cut blocks into smaller fragments
Blocks are too hard Soften blocks in detergent or phenol
Sections adhere to the knife or other parts of the machine Static electricity due to low atmospheric humidity Breath out or blow gently on the block and knife to break up static electricity or boil water in the room to increase the humidity
Knife edge is dirty Clean the knife edge
Knife edge is dull Sharpen the knife
Knife edge is too great Reduce the tilt
Ribbon is split or lengthwise vertical scratches are seen on sections Nicks are damage on the knife edge Sharpen the knife
Dirty embedding medium Re–embed in filtered wax
Knife edge is dirty Clean knife edge with xylene
Tilt of knife is too great Reduce tilt
Resistance is felt on the lower part of the section during cutting Tilt of knife is insufficient; paraffin block is therefore compressed against the base of the knife towards the end of stroke Increase the tilt
Horizontal or parallel lines or furrows across the section (chatters) are seen, forming thin and thick zones

Knife edge vibrates due to: (a) hardness of the tissue (b) tilt of the knife is too great

Treat with phenol during processing; or colloidionize Reduce the tilt
Section cut is sometimes thin, sometimes thick Knife is blunt Sharpen knife
Knife is not clamped properly Adjust the knife
Tilt of knife is too great Reduce the tilt
Knife or blockholder is loose Tighten adjusting and locking screw
Knife tilt is too small that block is compressed by bevel and section is not cut Increase the tilt
Knife makes a hard metallic scraping or ringing sound on backstroke, when section is cut Tilt of knife is too slanted or too big Readjust the angulation of the knife
Tissue is too hard Take fresh block treated with phenol during processing
Knife blade is too thin Charge the knife
Frozen tissue crumbles and comes off the blockholder when cut Freezing not adequate Refreeze the tissue block
Frozen tissue chips into fragments when cut Tissue is frozen too hard Warm the tissue with the fingers

Faults in Processing

What precautions should be taken for paraffin embedding?

Before delving into the specific techniques, these things should be considered with regards to general tissue embedding.

The paraffin of choice should have a similar hardness to that of the tissue being embedded. Melted paraffin should be kept 2-4 degrees Celsius above its melting point ; higher temperatures may separate the paraffin components, resulting in difficult ribboning.Cold plate temperature should be set between 3° C and -5º C. One cassette should be embedded at a time to prevent accidental transfer between cassettes. Forceps and molds should be clean and kept warm. Wiping forceps clean between specimens is advised.The grossing information should be referenced for the number of tissue pieces submitted as well as special instructions.Tissue should be embedded in the center of the mold, diagonally, and in an organized line (not randomly).The epidermis of all tissue pieces should face in the same direction in the mold, as shown in the drawing.Excisions, cyst cross-sections, and large tissue pieces should be embedded flat, with light pressure being applied over the tissue so that complete sections can be obtained during ribboning.Embedded tissue should be chilled rapidly on a cold plate to ensure strong paraffin support and adhesion around the tissue.

What are the most common laboratory safety problem?

E-Control Systems

Never store or consume food or drinks in labs where hazardous materials are used. This goes for keeping your lunch, snacks and sodas in refrigerators made for chemicals. Safety Eyewashes should be cleaned and flushed weekly. Always document the date of last maintenance on eyewash tags. Always wear appropriate personal protective equipment (PPE) when working in labs where hazardous materials are present. Never wear shorts or open-toed shoes in labs. Remove clutter and practice good housekeeping. Keep exits and aisles clear. Eliminate extension cords and power strips in series. Have the appropriate spill supplies available, and follow proper response procedures when spills occur. Maintain labels on chemical containers received from manufacturers and label secondary containers. Replace old and deteriorated labels. Segregate chemicals properly. Store acids in an acid cabinet or in a plastic container. Store nitric and other corrosive acids separately. Dispose of unwanted chemicals properly. Label chemical waste with specific contents. Keep waste tag attached to each specific waste container at all times. Keep chemical waste containers closed (do not forget to remove the funnel). Chemical fume hood sashes should be kept closed whenever possible. Maintain the minimum possible opening when working. Limit storage in hoods to essential items only. Always use a temperature monitoring system to ensure that volatile chemicals stay within their safe ranges. Also, items that will be used, such as vaccines, should be kept within their safe temperature ranges so they do not go bad. Always secure gas cylinders properly. Also always keep safety caps on gas cylinders when not in use.

: E-Control Systems

How can you prevent contamination in tissue culture laboratory?

How To Prevent Cell Culture Contamination Inside the body, cells are protected by the highly evolved and sophisticated immune system. Once isolated from tissues, cells in culture are “on their own”. We must therefore protect them with careful aseptic techniques to prevent their contamination from microorganisms that are present all around us, including inside the laboratory.

Various types of contamination exist: for example, cells can become contaminated with bacteria, mycoplasma, or mold. Some of them are harder to detect than others 1 (such as mycoplasma) and can therefore have a massive impact on your results without you knowing it. Others will be noticeable and will force you to interrupt your experiments – which wastes time, energy and money.

Contaminations in the laboratory can be a nightmare for scientists, especially as they can spread from one sample to all others present in the same incubator or hood. It is therefore an important responsibility for everyone in the laboratory to be as careful as possible, not only for the sake of their own experiments but for the sake of everyone else’s too.

  1. Measures to prevent contamination are numerous and should be applied throughout handling.
  2. With experience, these measures become second nature, but it can still be easy to make involuntary mistakes.
  3. While the field of cell culture is moving towards higher levels of automation 2 in part to prevent cell contamination, as of today we still rely mostly on manual labor, which presents many opportunities for mistakes in aseptic handling, especially for beginners.
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Here, we provide some essential tips to maintain an aseptic environment and prevent cell culture contamination.1. Wear gloves, lab-coats and use hoods This goes without saying, you must provide a barrier between your cells and the non-sterile environment that is your laboratory (or indeed yourself).

Using gloves, lab-coats and hoods does just that. Use your lab-coats only inside your cell culture lab and have them cleaned often. Make sure your hood is serviced often to guarantee that it is properly working. Note that there are different types of hoods, depending on the level of protection you want for the cells and the users 3,2.

Use your hood correctly Having a hood is not enough to ensure a sterile environment: you must use it correctly. First, make sure that you are not blocking the flow of air inside the hood: do not place material over the air outlets or inlets that are often found at the front and back of your bench space inside the hood.

In addition, make sure that you are working well inside the hood, and not too close to the edge to make sure that you are in fact operating under sterile conditions. You might want to empty your hood at the end of your day, and make sure to wipe it with 70% ethanol or 70% IMS before and after every use.

Some groups choose to have a waste disposal bin inside the hood, if you do, empty it and clean it often. Others prefer not to and keep it outside the hood as having used material or reagents for a long time inside the hood can increase the risk of contamination.

Finally, hoods often come with UV light: switch it on at the end of your day to sterilize your hood while it is not being used. Would you prefer to read this as a PDF? 3. Clean your incubator and water bath regularly Some new incubators have self-cleaning capacities – others must be cleaned manually. Do so often; the protocol will vary depending on the incubator.

If you use a tray of water inside the incubator, change the water often. You might sometimes need to add a chemical to the water to prevent contamination (always check that this chemical is compatible with the material your tray is made of). Similarly, the water in your water bath where you warm up your media should be changed regularly.

  1. Add a water bath treatment every time you change it.4.
  2. Spray EVERYTHING with ethanol or IMS Ethanol is typically used to kill bacteria.
  3. Make sure you use 70% ethanol mixed with water, and not a lower concentration.
  4. The water is known to increase the efficacy of ethanol in killing bacteria and some viruses.

Spray your gloves and everything that you bring inside the hood with ethanol. It is important to realize that every single time you touch something outside of the hood you should spray your gloves again before placing them back inside the hood. This applies to automatisms that might go unnoticed such as moving your chair, throwing something outside of the hood or touching your glasses.

  • Buy ethanol-proof markers to label your labware, otherwise spraying ethanol over it will remove your labeling.5.
  • Minimize exposure of cells to non-sterile environments Since so much effort is dedicated to maintaining a sterile environment inside the incubator and the hood, make sure that you minimize the time cells spend outside of these environments.

For example, when you check them under the microscope, or when you transfer them from the incubator to the hood. If you have to use your cells for extended periods of time in non-sterile conditions, for example during time-lapse imaging, and need to put the cells back in an incubator, designate a separate incubator for this purpose if possible.

  • This way, the increased contamination risk will be confined to one incubator.6.
  • Buy sterile reagents and keep them sterile! Be sure to buy reagents that are sterile.
  • This encompasses media, PBS or any other product that will come into contact with the cells.
  • If you use reagents for other work that is not sterile, such as antibodies, do not use them for your cell culture.

Keep two separate stocks of sterile and non-sterile reagents. In addition, you can also choose to filter your media (or any other liquid reagent) through membranes that remove bacteria. Typically, 0.2 μm filters are enough to block bacteria. Another way to maintain the sterility of your reagents is to aliquot them.

That way, if one aliquot gets contaminated the entire stock is not affected, and you can throw away the one aliquot instead of the entire stock. You can also buy pre-aliquoted reagents.7. Use sterile labware Apart from reagents, you must also sterilize the labware that cells will be in contact with. To sterilize these, you can use an autoclave machine.

Make sure however that your labware can be autoclaved! Not all materials can sustain the temperatures of the autoclave. For example, scientists often autoclave pipettors, or pipette tips. If you can, buy consumables that are already sterile, such as flasks.

To maintain sterility, open the bag of flasks only once you have sprayed it and placed it inside the hood. When finished, close the bag inside the hood before bringing it back outside. If you seal the bag with tape, and the tape is outside the hood, keep the bag closed with your hands until you tape it.

You can also autoclave water, if you have a source of filtered water in your laboratory. Place a pressure-sensitive tape on the material before placing it in the autoclave: if the material has successfully been autoclaved and sterilized, the tape should change color.8.

  1. Use filter tips and change them often Pipette tips that contain filters prevent the solution inside the tip from touching the pipettor.
  2. Without filter tips, your pipettor can become contaminated, thus contaminating every other sample that it touches First, this means that if one sample is contaminated with microorganisms these can spread to other samples, but it can also mean that you can introduce cells involuntarily from one sample to the other.

For the same reason, make sure you change your tips often during your experiments, especially between samples. When you operate with the pipettor be careful as to not bring the pipette tips in direct contact with anything else, and if you do change tip.

Importantly, if you want to pipette something out of a bottle, avoid using a pipettor that you must insert inside the bottle, to prevent the pipettor from directly touching the inside of the bottle: instead, use a pipette gun that can be used with longer pipettes that can reach deeper inside the bottle.9.

Check your cells often Even though some contamination, such as that associated with mycoplasma, is hard to detect visually, it is always wise to check your cells regularly under the microscope. If you do, try not to keep your cells away from the incubator for too long to minimize the risk of contamination.

  1. You can check for moving bacteria (do not confuse with cellular debris!), morphological changes or unusual proliferation rates.
  2. If you suspect your sample is contaminated with mycoplasma, there are commercial kits available to confirm this.10.
  3. Bleach your contaminated samples If you do think your sample is contaminated, fill it with 10% bleach, Trigene or Chloros in the sink or away from the incubator for a while.

Empty it inside the sink and throw away the culture vessel. If you realize that your sample is contaminated, warn your labmates, especially those who have samples in the same incubator! This is really important, it’ll allow them to quickly react, check that their samples are fine and plan accordingly if they are not.

  1. If it’s possible to empty the incubator of all its samples, clean the incubator.11.
  2. Use good labeling practice Keeping good records of your stocks of cell supplements and media will help you identify sources of contamination more quickly.
  3. If you find that some cells are contaminated, you should be able to identify the stock to which these cells belong to and test whether the contamination is common to the entire stock.

Similarly, if you know the batch of media or supplements you were using, you can check or throw away the affected stock to avoid repeated contamination. Good labeling can also prevent contamination of your experiments with other cell types: you want to make sure the cells you have inside your assay are the ones you think they are.

  1. This is a huge problem in the field, with up to 30,000 studies using the wrong cell lines 4,
  2. Therefore, make sure you label all your flasks and vials properly, to minimize the possibility of cross-contamination between samples.12.
  3. Common sense Last but not least, common sense goes a long way to prevent contamination.

For example, do NOT perform work with bacteria or molds in your cell culture lab. Needless to say, no sample containing bacteria should be placed in your cell culture incubator. If you are ill, try to refrain from doing cell culture work (we appreciate this may not always be possible).

  • Minimize talking in front of open incubators or whilst working at the hood.
  • Do not leave incubator doors open for too long.
  • Wash your hands before and after cell culture handling! References 1.
  • Nikfarjam, L.
  • Farzaneh, P.
  • Prevention and detection of Mycoplasma contamination in cell culture.
  • Cell J,13, 203–12 (2012).2.
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Wendt, D., Riboldi, S.A., Cioffi, M. & Martin, I. Potential and bottlenecks of bioreactors in 3D cell culture and tissue manufacturing. Adv. Mater,21, 3352–3367 (2009).3. Cell culture contamination – Thermo Scientific.4. Horbach, S.P.J.M. & Halffman, W. The ghosts of HeLa: How cell line misidentification contaminates the scientific literature.

What is the common contamination in tissue culture?

Bacteria and fungi – Bacteria, molds, and yeasts are present virtually everywhere and are able to quickly colonize and flourish in the rich and relatively favorable environment available for tissue cultures. These microbes are the most commonly encountered cell culture contaminants, because of their size and fast growth rates.

  • We can easily detect these microbes in cultures within few days when we don’t supply antibiotics to the culture medium.
  • The best way to keep your plants free of microbial contamination is to perform daily observations.
  • This practice can help you to protect your cultures in the long run.
  • However, if you regularly supply antibiotics to the media, then these organisms can develop resistance and convert into slow-growing or low infection-causing organisms.

Eventually, they become very difficult to detect just by visual observations. As a result, many laboratories use microscopes for the identification of these microbial cells.

What are the risks of tissue culture?

While tissue culture presents numerous benefits, it is not exempt from certain limitations. –

Tissue culture can require more labor and cost more money in building the facility and equipping the lab with all the instruments and chemicals. There is a chance that the propagated plants will be less resilient to diseases when grown in outside conditions due to the type of environment they are grown in. It is imperative that, before being cultured, the material is screened; failure to pick up any abnormalities could lead to the new plants being infected. While the success rate is high if the correct procedures are followed, success with the tissue culture is not a guarantee. That’s why accurate protocols are necessary to grow plants in tissue culture setting, which can be laborious when you try to create one working protocol by yourself. Contamination is the major issue in tissue culture setting. Plants can get infected by bacteria, fungi, and viruses. That’s why all measures should be taken and PPE kit should be used while performing tissue culture in your lab. Tissue culture is an advanced technique and require some advanced knowledge and practice for anyone to get started in the field.

As you can see, the advantages do seem to outweigh the disadvantages. Sure, you may have to spend a bit more money to get your DIY tissue culture going, but the rewards certainly outweigh the initial cost. So, let’s take a look at the Tissue Culture Process and see if we can break down the complicated terms into something a little more digestible. Types of Tissue Culture Technique Tissue culture is a technique in which healthy tissues are extracted from living matter or organisms. In plant tissue culture, this could be either the leaves or other parts of the plant- depending on the protocol. Based on the explant (starting material/plant tissue used to grow plants), tissue culture is classified into the following types :

Callus Culture: A callus refers to a cluster of undifferentiated cells with the remarkable ability to give rise to various plant parts. When the plant tissues derived from any plant organ are artificially induced in laboratory settings, they form callus, which further give rise to different plant organs, roots and shoots. Seed Culture: Protoplast Culture: A protoplast is a plant cell lacking a cell wall. In this technique, the cell wall of plant cells is eliminated through mechanical or enzymatic means. The resulting protoplasts are purified and subsequently, under controlled conditions, the cell wall is regenerated before transferring them to appropriate media for further growth into a complete plant. Meristem Culture: Meristem culture involves the isolation of the meristematic region, such as shoot tips, from plants and its transfer to a growth medium containing nutrients, vitamins, and plant hormones. This technique promotes cell division and tissue differentiation in the cultured cells. Meristem culture finds diverse applications, including the production of disease-free plants, regeneration of complete plants, generation of transgenic and haploid plants, crop enhancement, and preservation of germplasm. Embryo Culture: Embryo culture involves the isolation and cultivation of either immature or mature plant embryos to support their development into complete plants. Instead of individually sterilizing the embryos, this method involves sterilizing the organ (such as ovule, seed, or fruit) from which the embryos are derived and utilizing it in the culture process. Ovary Culture: The technique involves culturing fertilized or unfertilized ovaries of plant species in a suitable environment to facilitate their development into complete plants. This method, also known as gynogenesis, is primarily employed to overcome barriers before and after fertilization. Additionally, it has been utilized to achieve interspecific hybrids. Anther/Pollen culture: Pollen/anther culture is a technique in plant biotechnology where pollen grains or anthers (the male reproductive parts of flowers) are isolated and cultured in a nutrient-rich medium. This method allows for the development and regeneration of haploid plants or callus tissues from the cultured pollen or anther cells. It is commonly used in plant breeding and genetic studies to produce new plant varieties or to study the behavior of plant cells in a controlled environment.

What Make Tissue Culture So Great? Tissue culture can be used in the reproduction of a wide variety of species and has many practical applications. By using the tissue culture process, a plant’s yield can be increased dramatically, and in a short amount of time.

  • The plant can also be genetically altered so that it becomes immune to certain diseases and viruses.
  • The genetic modification enables growers to ensure that plants carry very specific characteristics.
  • In many cases, businesses and individuals will propagate the plants to carry specific traits that are more profitable for their business, or more desirable for personal use.

On another note, the tissue culture process can be used to promote the survival of a rare plant or endangered species. Lastly, the tissue culture technique relies on the plant’s innate ability to rejuvenate cells quickly, and these rejuvenated cells are copies most often referred to as clones.

This technique can be used in a lab with expensive and complicated equipment, or it can be simply adapted for a home DIY. Plant Cell Technology is a leading provider of tissue culture products and services. It offers tissue culturists like you all the products you need to perform your in vitro experiments, ranging from MS media, agar, gellan gum, and culture vessels, to hormones and our proprietary product plant preservative mixture (PPM).

Further, if you’re stuck at any stage of the tissue culture process, you can use our consultation services to have a video chat with our tissue culture expert. He will give you an instant potential solution to your problems to keep your lab experiments running! And, if you are new to the area and interested in growing your plant business using this amazing technology or want to learn this propagation technique as a hobbyist, you can join our tissue culture master class, The classes are curated to turn you into a tissue culture pro.

What are the specific hazards in the clinical laboratory?

Common Clinical Laboratory Hazards and Waste Disposal It is very important to protect laboratory worker from hazards. The laboratory hazards fall into three main categories: chemical hazards, biological hazards, and physical hazards. The major chemical hazards in laboratories are the cleaning agents, anesthetic gases, disinfectants, drugs, and solvents.

The persons working in laboratory are exposed to chemical hazards during their usage or due to improper storage. Biological hazards involve exposures to infectious samples, animal diseases transmissible to humans, and biological agents used during experimental procedures that include viral vectors, etc.

The exposure to physical hazards is associated with research facilities. The laboratory personnel encounter the physical hazards due to accidental spill of corrosive reagent, broken glassware, etc.

What is hazardous in safety?

What are hazards? – A hazard is a source or a situation with the potential for harm in terms of human injury or ill-health, damage to property, damage to the environment, or a combination of these. Hazards at work may include noisy machinery, a moving forklift, chemicals, electricity, working at heights, a repetitive job, or inappropriate behaviour that adversely affects a worker’s safety and health.

What are MSDS safety hazards?

What is a Material Safety Data Sheet (MSDS)? – Back to top A Material Safety Data Sheet (MSDS) is a document that contains information on the potential hazards (health, fire, reactivity and environmental) and how to work safely with the chemical product.

It is an essential starting point for the development of a complete health and safety program. It also contains information on the use, storage, handling and emergency procedures all related to the hazards of the material. The MSDS contains much more information about the material than the label. MSDSs are prepared by the supplier or manufacturer of the material.

It is intended to tell what the hazards of the product are, how to use the product safely, what to expect if the recommendations are not followed, what to do if accidents occur, how to recognize symptoms of overexposure, and what to do if such incidents occur.

What is risk assessment of hazards in laboratory?

Laboratory Safety A risk assessment should identify potential hazards and determine controls that can be implemented to eliminate or reduce any risks to employees, operations, and/or property. A new risk assessment should be completed and documented each time a new potentially hazardous experiment is carried out.