Colony Counter Compatibility Mode
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The document discusses colony counters, which are instruments used to count bacterial colonies growing on agar plates. Early colony counters involved manually marking colonies on plates placed on a lighted surface and counting by hand. More recent automatic colony counters can electronically identify and count colonies by detecting contrasting light and dark spots, and are connected to computers to further analyze and magnify individual colonies. The recent colony counters provide faster, easier, and more efficient counting and observation compared to older manual methods.
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2) Injecting India ink under the embryo to aid visualization.
3) Lowering dye-filled pipettes using a micromanipulator to inject the embryo in specific rhombomeres.
4) Sealing the window and incubating the eggs until sacrifice and analysis. Proper injection placement can be ensured by early dissection and sectioning.
Isolating Chromosomes
This document describes the process of isolating and preparing chromosomes from white mice for analysis. Key steps included treating mice with colchicine to arrest cells in mitosis, sacrificing the mice and extracting bone marrow cells and testes tissue, suspending the cells in a hypotonic solution to spread the chromosomes, fixing and staining slides with the cell suspensions, and observing the chromosomes under a microscope. The white mice were used as model organisms because they are readily available, low-cost, easy to handle, and their chromosomes correlate well with other test organisms.
Autoclave
1. An autoclave is a device that sterilizes equipment and supplies by subjecting them to high pressure steam at 121°C for 15-20 minutes.
2. When autoclaving glassware, cap Pyrex bottles loosely and cover non-safety glass bottles with aluminum foil to prevent explosions.
3. Proper autoclave use involves following safety procedures like wearing protective equipment, waiting for full pressure release before opening, and allowing time for contents to cool before removal.
Aseptic Technique
Aseptic Technique
The media on which you culture desirable microorganisms will readily grow undesirable contaminants, especially molds and other types of fungus, and bacteria from your skin and hair. It is therefore essential that you protect your cultures from contamination from airborne spores and living microorganisms, surface contaminants that may be on your instruments, and from skin contact.
Bacteria and other contaminants cannot fly. Nearly all forms of contamination are carried on microscopic dust particles that make their way onto sterile surfaces when they are carelessly handled. One exception is insect contamination, such as by ants for fruit flies. Fruit flies are a particular nuisance because they can crawl under the lids of agar plates and lay eggs. You would think that people doing genetics research would have developed a model by now that can't fly into other peoples' experiments!
A contaminated culture can often be rescued, however there is always the risk that you will re-isolate the wrong microorganism. Besides, you don't have that kind of time to waste. Exercise extreme care to keep your cultures pure.
Agar Culture Medium
To prepare an agar culture medium, one must first sterilize empty petri dishes by boiling them and allowing them to dry upside down. Agar powder is then prepared by mixing it with water in a microwave according to standard directions. Bottled agar formula is also warmed in the microwave with the cap loosened but not removed. Once the agar is liquid, petri dish lids are cracked open and the agar is carefully poured into the dishes under the lid to prevent contamination, setting to a thickness of 1/8 inch before storing upside down once solidified.
Allium Cepa Genotoxicity Test
Onion (Allium Cepa) Genotoxicity Test
Laboratory of Ecotoxicology and LCA
Department of Environmental Chemistry, ICT Prague
References:
1. FERETTI, D., ZERBINI, I., ZANI, C., CERETTI, E., MORETTI,M.,MONARCA, S. (2007): Allium cepa chromosome
abberation and micronucleus tests applied to study genotoxicity of extracts from pesticide-treated vegetables and
grapes. Food Addit. Contam. 24 (6): 561-572.
2. RANK, J., NIELSEN, M.H. (1997): Allium anaphase-telophase genotoxicity assay. Department of Environment,
Technology and Social Studies, Roskilde University, Denmark.
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- 3. Colony counter • A colony counter is an instrument used to count colonies of bacteria or other microorganisms growing on an agar plate. • An instrument designed to conveniently count or assist counting colonies of microorganisms on a plate containing a gelled growth medium.
- 4. Colony counter • One variety uses a pencil-like rod with a metal tip, which is connected by an electrical connection to the gelled growth medium; when touched to a colony on the plate, the completion of the electrical circuit causes an increment of 1 unit on the readout of the colony counter.
- 5. Early counters Recent counters • Early counters were merely lighted surfaces on which the plate was placed, with the colonies marked off with a felttipped pen on the outer surface of the plate while the operator kept the count manually. • More recent counters attempt to count the colonies electronically, by identifying individual areas of dark and light according to automatic or user-set thresholds, and counting the resulting contrasting spots.
- 6. (for the automatic colony counter)
- 7. 1. Place the Petri-dish on the colony counter 2. Count a Petri-dish by clicking the count button on the software.
- 8. • 3.The results will be displayed indicating the number of colonies.
- 9. The Recent Colony Counter • The automatic colony counter has the ability to separate the image of combined colonies for the user to magnify each colony. • The recent colony counter is connected to a PC.
- 10. For the old/digital colony counter
- 11. The Old Colony Counter 1. Place the colony counter on the lighted surface with the colonies marked off with a felt-tipped pen on the outer surface. 2. Keep the count manually
- 12. Comparison Recent Old • Is indeed faster and easier to use • More efficient for observation. • Hard to use • Manual counting • Not so efficient compare to the recent