ZOO101 Lab Manual Complete

CONTENTS · Laboratory Safety · Practical’s:

1. Preparation of Slides (Dry Mount)
2. Preparation of Slides (Wet Mount)
3. Observation of Wet Mounts of Human Cheek Cells Employing Bright and Dark Field Microscopy
4. Measurement Cell Size: Prokaryotic (Bacterial Cell) Cell Size.
5. Measurement of cell size: Eukaryotic (Human Cheek Cell) Cell size.
6. Recording of Microscopic Observations with the Help of Camera Lucid
7. Liquid Handling: Proper Use of Pipettes.
8. Liquid Handling: Proper of Micropipettes.
9. Hematoxylin and Eosin Staining (Harris’ Hematoxylin and Eosin).
10. Gram’s Staining.
11. Handling of Centrifuge Machines.
12. Paper Chromatography of Amino Acids.
13. Thin Layer Chromatography of Amino Acids.
14. Spectrophotometric Estimation of Glucose.
15. Preservation of Representative Animals of Various Phyla(vertebrates)
16. Preservation of Representative Animals of Various Phyla (Invertebrates) Molecular biology techniques are applicable not only to modern medical practice but also to the identification of genetically modified organisms, forensics, and quality assessment of laboratory animals, pharmacogenomics, and other fields. This procedure manual encompasses most widely used molecular biology techniques namely DNA extraction, end-point and Real-Time PCR.

Safety Considerations:

•  Use personal protective equipment such as disposable gloves, lab coats, disposable masks, etc.
•  Handle all sharps with care and dispose of sharps in the sharp’s disposal containers.
•  Handle hazardous chemicals and samples carefully. Blood and other body fluid must be considered potentially hazardous. Biological waste should be disposed of in the designated trash bags that could be incinerated later on.
•  Decontaminate the work benches regularly and especially before and after work. For decontamination, wipe the surfaces with 10% bleach followed by water.

Minimum Standards and Controls:

•  During DNA extraction, PCR and other experiments appropriate standards and controls should be used to assure the quality of the results.
•  Positive control to check the efficiency of the reagents, procedure and equipment.
•  Negative Control to check the contamination

Validation of Critical Reagents and Procedures:

All technical procedures and critical reagents should be tested and validated before performing the actual case work or research work experiments.

Calibration of Instruments:

All instruments should be calibrated according to required schedule and before performing the validation studies, case work and research experiments

Practical 01:

Making microscopic slides is a great skill. The first step in making Microscopic slides is to choose the type of mount. The "mount" is simply the way in which a specimen is placed on the slide.

When we are making microscopic slides there are three types of mounts we can Prepare. These are wet mount, dry mount, and prepared mount. Each mount has its own Pros and cons.

Dry Mount Microscope Slides:

Dry mounts microscope slides are the easiest to make.

Material:

•  Glass slide Cover slips.

Dry mounts work best for samples like:

•  Pollen
•  Hair
•  Feathers
•  Or even dust particles caught in a microfilm filter.

Steps to Make Dry mount Slides:

The basic steps to Make dry mount slides are very simple. Place the sample on the slide and cover. The cover will protect both your sample and lenses (Objective).

Note:

These mounts are temporary unless you seal the cover slip in some way. It is harder to see the more complex structures of some species. For that reason, you will have to learn how to make a prepared mount.