Cell And Molecular Biology Lab

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Circular piece of DNA sperate DNA to cell

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Plasmid

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Important for replication, transferability and Resistance

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Plasmid

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Found on the certain parts of the jellyfish to ability of light will be the market or reported protien

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GFP

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Emits light in use in monitoring movement of the protein behavior

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GFP

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Calcium indicators

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Calmodium

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It helps us to understand signaling activity

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GFP florescence

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Target gene promoter+ GFP sequence

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Signaling promoter

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Benefits of florescence labeling

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1. Labeling and tracking protein localization

2. Labeling subcellular compartment

3. Checking strength of the promoters

4. Dynamic process like vusicular

5. Studying cell signaling promoter

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Knowing the tool and the guidelines on the lab, putential danger, rid of the guidelines

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Labsafety

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Labsafety remembered

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1.Know the rules the guidelines, potential danger

2. Detect and removal of the potential risk

3. Preparation for emergency action

4. Lucy

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A handling material and an infectiousness agent contaminated principles, method, procedure

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Biosafety

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No containment, unlikely to cause disease

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BL-1

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Have containment, moderate risk varying severity

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BL2

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High containment, aerosol and has potential lethal

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BL3

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Max containment, exotic and life threatening

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BL4

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Steps in Biosafety procedure

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1.Sample collection

2. Choosing right laboratory

3. Proper ventilation

4. PPE

5. Conduct aseptic technique, sterility test

6. Incubation

7. Result and decontamination called fumigation

8. Disposal of the use material and autoclaving

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Indispensable tool for biological research in able research to visualize structure with detail

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Microscopy

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Formula of total magnification

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Total magnification= objective mag and eyepiece

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Vital in microscopy is a preparation of specimens in the stage

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Mounting

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Staining step

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1. Dehydration

2. Permealization

3. Fixation

4. Mordant application

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Replacing water of a chemicals compounds

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Dehydration

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Dissolving cell membrane to penetrate dye

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Permealization

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Chemical bond between protein to increase rigidity

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Fixation

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Strong complex dye and tissue

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Mordant application

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Some staining method

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Mallory method

Hematoxylin and Eosin staining

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Sample preparation on microscopy

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1. Collection

2. Fixation

3. Embedding

4. Sectioning

5. Staining

6. Observing

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In Simple microscopy remember

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1. Assembling a microscope part

2. Staining method desicion

3. Mounting and find magnification

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Use of flourescent chemical compound that re-emit hight excitement

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Flourescent microscopy

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Keeping the cell the composition

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Fixation

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Why we use immersion oil

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Coz oil reduces refraction of light to see clearer and crispier views of the specimens

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Use nanoscale visualization use beam that cause no storage of col5

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Electron microscope

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Permits to analysis of cell morphology and abundance

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Staining

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Camera or monitor for getting image

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Micrograph

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Capture image both fixed and ectopic activity of the cell

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Confocal microscopy

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Allowed us to capture collection of image render to assist what has the infection

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3D rendering

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Use in confocal that use Flourophore

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Green PMT

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Confocal microscopy uses autoflourecense

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Red PMT

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Increase the background noise regulation

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Scanline

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Slice the specimens into 3D image

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Z- axis

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Light traveling in confocal microscopy

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1. Laser lights

2. Dicroic mirror or beam splitter

3. Detector

4. Objective

5. Specimens

6. Pinhole

7. Camera

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Know the morphology it will be known the species of the infection and prepare what possible action to resolve

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Confocal microscopy

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Cuts the gene of the interests

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Restriction enzyme

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Help to bind the plasmid and gene of interest (glue)

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DNA ligase

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Induce GFP to florescence

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Calcium binding

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GFP bind with?

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Calmodium with sequence light chain