What are five safety factors to consider when working with bacteria?

What are five safety factors to consider when working with bacteria?

Microorganisms Safety Guide

• Treat all microorganisms as potential pathogens.
• Sterilize equipment and materials.
• Disinfect work areas before and after use.
• Wash your hands.
• Never pipette by mouth.
• Do not eat or drink in the lab, nor store food in areas where microorganisms are stored.
• Label everything clearly.

Is microbial culture dangerous?

Culturing microorganisms can lead to the growth of dangerous pathogens. Pathogens may enter the human body through skin, eyes, puncture wounds, inhalation, or ingestion. Students who have compromised immune systems should consult the teacher or their doctor before participating in microbiology experiments.

Why bacteria is used in laboratory culture?

Microbial cultures are used to determine the type of organism, its abundance in the sample being tested, or both. It is one of the primary diagnostic methods of microbiology and used as a tool to determine the cause of infectious disease by letting the agent multiply in a predetermined medium.

What are the three main types of microbiological culture media?

Depending upon the addition and quantity of this substance, media are of three types:

• 3.3.1 Liquid (Broth) Media. Liquid (Broth) Media, such as nutrient broth, tryptic soy broth or glucose broth which are prepared without the use of agar agar.
• 3.3. 2 Semisolid Media.
• 3.3. 3 Solid Media.

What precautions should be taken during Subculturing?

Care should be exercised in the opening of tubes with tight caps to prevent the breakage of the glass. Care should be taken to avoid contact with skin, eyes, or mucous membranes when handling culture media or any laboratory reagent, stain, fixative, or chemical. If contact occurs, flush immediately with running water.

What is the main purpose of Subculturing?

Subculturing prolongs the lifespan of the cells or microorganisms, allowing for long-term maintenance and observation of the culture. The process of subculturing involves transferring microbes from one growth container to another, providing the microbes with a fresh supply of nutrients on a solid or liquid medium.

What is the purpose of Subculturing?

What is the purpose of subculturing? Subculturing keeps cells and microorganisms alive by transferring them from a previous growth culture to a fresh growth medium that allows for increased growth and multiplication.

How do you maintain stock culture?

Stock Culture Maintenance and Storage

1. Effective maintenance of stock cultures is essential for QC, method validation and research purposes.
2. Repeated subculturing may eventually lead to contamination, loss of viability and genotypic/phenotypic changes.
3. Freeze-drying and cryogenic storage are preferred, but may not be practical for smaller laboratories.

Which stock culture are used frequently?

working stock cultures

How do you prepare a bacterial stock culture?

After you have bacterial growth, add 500 μL of the overnight culture to 500 μL of 50% glycerol in a 2 mL screw top tube or cryovial and gently mix. Note: Make the 50% glycerol solution by diluting 100% glycerol in dH20. Note: Snap top tubes are not recommended as they can open unexpectedly at -80°C.

How do you revive ATCC culture?

For freeze dried cultures, using a single tube of the recommended media (5 to 6 mL), withdraw approximately 0.5 to 1.0 mL with a Pasteur or 1.0 mL pipette. Use this to rehydrate the entire pellet, and transfer the entire suspension back into the broth tube and mix well.

How do you use ATCC culture?

Initiating Lyophilized Cultures Using a Pasteur pipette, aseptically add 0.5 mL of the recommended growth medium to the freeze- dried material. Mix well. 2. Transfer the entire suspension to a test tube containing 5 to 6 mL of the recommended medium.

Why must thawing of cells be done quickly?

It is important to thaw rapidly to minimize any damage to the cell membranes.

Does freezing kill cells?

Freezing usually damages cells because water expands when it freezes. Animal cells just have thin membranes around them. When ice crystals form, they destroy the cells. That’s what frostbite is.

How do you recover cryopreserved cells?

The recovery of cryopreserved cells is straightforward: Cells are thawed rapidly in a water bath at 37°C, removed from the freeze-medium by gentle centrifugation and/or diluted with growth medium, and seeded in a culture vessel in complete growth medium.

Should cells be thawed slowly?

Most recent answer. For the 1st question, a slow cooling rate is generally needed for mammalian cells to avoid the lethal intracellular ice formation (IIF). The optimal cooling rate should be slow enough to avoid IIF and fast enough to avoid severe dehydration. Different cell types have different optimal cooling rates.

How do you revive cells?

Thaw frozen cells rapidly (< 1 minute) in a 37°C water bath. Dilute the thawed cells slowly before you incubate them, using pre-warmed growth medium. Plate thawed cells at high density to optimize recovery. Always use proper aseptic technique and work in a laminar flow hood.

How do you restore cells?

Guidelines for thawing cells

1. Thaw frozen cells rapidly (< 1 minute) in a 37°C water bath.
2. Dilute the thawed cells slowly before you incubate them, using pre-warmed growth medium.
3. Plate thawed cells at high density to optimize recovery.
4. Always use proper aseptic technique and work in a laminar flow hood.

How do you bring up the cells in frozen?

Video: Thawing cells

1. Remove the cryovial containing the frozen cells from liquid nitrogen storage and immediately place it into a 37°C water bath.
2. Quickly thaw the cells (< 1 minute) by gently swirling the vial in the 37°C water bath until there is just a small bit of ice left in the vial.

How do you preserve cell lines?

The only effective means of preservation of animal cells is by freezing, which can be accomplished with either liquid nitrogen or by employing cryogenic freezers. The freezing process involves slowly reducing the temperature of prepared cells to -30 to -60°C followed by a transfer to temperatures less than -130°C.

How do you thaw a cell line?

Thawing Method

1. Remove the frozen vial from the liquid N2 storage.
2. Loosen the cap of the vial slightly to release the pressure inside.
3. Thaw the cells in a 37ºC water bath.
4. Keep the lid of the freezing vial above the surface of the water to lessen the chances of contamination.

What is cell passaging?

Subculturing, also referred to as passaging cells, is the removal of the medium and transfer of cells from a previous culture into fresh growth medium, a procedure that enables the further propagation of the cell line or cell strain.

How do I start my own cell line?

The simplest way to create a new cell line is to modify an existing one, a common strategy when an established line already comes close to meeting the requirements. Cells optimized to grow particular viruses or maximize recombinant protein production often come from such modifications.

What are the different types of cell lines?

Attached cell lines can be classified as 1) endothelial such as BAE-1, 2) epithelial such as HeLa, 3) neuronal such as SH-SY5Y, or 4) fibroblast such as MRC-5….Cell Morphology Types.

Attached Cell Lines
Jurkat	Human T-cell Leukemia	Lymphoblast
THP-1	Human Monocyte Leukemia	Lymphoblast