hystology tec.

List in odern the methods for optical micropcopy

( federico deve essere sempre solo)

1. FIXATION to isolate thetiddu ex. Woth formladheide or methanol glutnaldheide for EM)

2. Dehydration so that you can use a non polar solvent

3. EMBEDDING to make it fixed ex. In wax

4. SECTIONING ex. Microtome or cryosectioning

5. Staining so that you have contrast

What is the function of fixation, what. Do you use?

What about dehydration what is the differenc eif you have to use EM?

Fixation to preserve the structure

Use ethaonleexc. Gtuanladheide for EM

The difference is that for optic microscope you use PARRAFINE to denaturate proteins

When is section not necessary? What is the purpose?

So thta the light can go though

Microtome or criosection

AS THEY ARE SUSPENSIONS IN BLOOD AND BONE MARROW IS NOT NECESSRAY

STAINING. What do i use for the cytoplasm? What for nucleus, exc

The cytoplasm is basic so ELECTROPHILIC use EOSIN - ROSSA

The nucleus is acidic so BASOPHILIC use HEMATOXIUM + BLU

When do i use negative stain EOSIN

For eletrophilic specific so Basic so cytoplasm

When do i use hematoxium + blu

Anyting that is acidic

What stain are used for glycoprotein and polysaccarides?

What are used for the ecm?

What is used for lipids

Glycoprotien: PAS

Ecm: TRICHROME

LIPID: red oil and sudan black

When do i use PAS, trichorme?

Pas: glycoproteins and polysaccaride

Trichrom : ecm

What is the differenc ebtween direct and indirect immunofluoresnce?

In the direct you use ANTIBODY VS ANTIGEN but is LOW SENSITIVTY

Indirect you use MARKED COUPLE ANTIBODY VS ANTIBODY so it more sensitivity!!

Fluorescence micorpscope is based on ?

Different asborption of wavelenght

Confocal microscopy, what utilizes? What allows to make as an image

By focusing the light on one plan and removing the light above nad bleow and with LASER youu hav e a3d image

Resolution power!!

Optic,

Tem

Sem

Afm

Optic: 0.2micrometr

Tem is 0.05-1 nm

Sem 2,5 nm

Am 50 picom

If i want just to see the TISSUE MORPHOLOGY what can i use

Optic micropscopy for tissue morphology

Which allows you to see in 2d organlles INSIDE

Tem

Tem is 0.05 1 mm of resoloution

2d image

Very high resolution

What you cn see the 3D and the surface visualization’

SEM

SURFACE, HIGHER DEPTH PF FILED, LESS RESOLUTION OF TEM , 3D

AFM 2 main purpose

1. Mechanical properties at the structural levl

2. Topography

Which tequinique is to put the tissue in a organic solvent

Dehydration

What tecquine is based on the anionic, cationic properties to stain it

Hemoxin and eosin

Glycogen would be bast stain from

Pas as it is a polysaccaride

What microsocpe need HEAVY METAL COMPOUND to fix

TEM

Best thing to use for a protein in the ecm

Immunoistochemistry

What can you visualize with in situ hybridization

Nucleic acids