MICROSCOPY AND STAINING

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ALGAE-spirogyra

look for chloroplast with green pigments, look for the nucleus

PROTOZOA- amoeba

look for the nucleus, amoeba crawl using pseudopods

YEAST- S. cerevisiae

These are unicellular fungi, look for the smaller pieces attached to the cells called buds (this is a type of asexual reproduction)

MOLDS- Aspergillus

these are multicellular fungi, look for the filamentous cells called hyphae, look for spores

PRIMARY STAIN

stains all cell the stain

MORDANT

enhances the primary stain and allows better penetration of stain into certain cell types

DECOLORIZER

removes color from some cells while leaving it in others

COUNTERSTAIN

adds a second round of stain to all cells

GRAM STAIN

determines type of cell wall

ACID FAST STAIN

specific for Mycobacterium

ENDOSPORE STAIN

specific for endospores in Bacillius and Clostridium

SIMPLE STAIN

capsule stain- Identifies bacteria with a capsule. Bacteria with capsules are identified as pathogens (they will cause diseases).
Example: Streptococcus and Pseudomonas

PREPARING SLIDES
method1: Wet mount technique

add sample in a drop of water to the slide
add stain if needed
cover with coverslip

PREPARING SLIPED
method2;

add sample in a drop of water to a slide
allow to air dry
heat fix by passing several times quickly through bunsen burner flame
-heat fixing attaches to cells to slide, kills the cell, open cell wall to allow stain to enter.
add stains after heat fixing

01.When doing a gram stain procedure, what color is a gram-negative cell after decolorized has been added ?

The gram- negative cell is always pink

02: In this diagram, why is the decolorized not forcing the color from the cell?

The cell wall is too thick and has no priors to allow the larger CV-I complex out

03: From this diagram, what is the thick, yellow layer surrounding the cell?

The thick, yellow layer is the mycelia acid layer.

04:In the acid fast staining procedure, why does the methylene Blue not add to the cell’s layers and create a purple color?

all options are correct

05: In the Spore staining, what is the mordant in the procedure

Heat

06:In the spore staining, why does the spore not stain red like the cell?

the spore has many layers of protection that keep chemicals out

07: what genus of bacteria can be identified by a spore stain procedure?

Bacillus or Clostridium

08: If we identify spore on a spore stain slide, what type of environment were the cell in before the spore stain procedure to see these results

The cell were in an unforvarable environment to create spore

09:If we do a spore stain procedure and we see circular red cell, what do we know about these cells?

circular cells will not make spore because they are not Bacillus or Clostridium

10: what do we know about the cells that stain pink in a gram stain procedure

we will know their cell wall is negative, we could also see shape and arrangement

11.Looking at this diagram, what type of staining was done here?

capsule stain

12: when you look math this stained slide(blue and pink), what type of staining is this?

Acid fast stain

13: When you look at this stained slide(blue and pink), what disease might be associated with this slide?

this may show us TB or Leprosy

14: When you look at this stained slide(green and red), what type of staining is this?

Spore Stain

15: From this stained slide (green and red), what diseases might be associated with this slide?

This may show Anthrax or tetanus