Staining for Conventional Light Microscopy

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Transcript of Staining for Conventional Light Microscopy

Stain

StainingStain

Noun Verb

Noun

What is stain and staining?

Unstained tissue Stained tissue

Why do we stain?

Why do we stain?

Dr Farhana Taher Sumya Guided by: Prof Khondkar Manzare Shamim

Dept. of Anatomy, BSMMU

STAINING For Conventional Light Microscopy

What should the audience be a able to do after attending this session?

1. Define stain and staining

2. Explain what is achieved with staining

3. Explain how staining for conventional light microscopy take place

4. Explain some basic terms on staining

What should the audience be a able to do?

5. Explain the causes and solutions of the problems commonly faced in staining

6. Cite examples of specific tissue components stained with special stain

Staining Technique

Vital

Supravital

Intravital

Nonvital

How can we stain?

Supravital staining: the cells with the dark blue dots and curved linear structures

(reticulum) are reticulocytes.

How can we stain?

Nonvital staining

How can we stain?

T

T ST

T

T

S

M S

SA

How can we stain?

T

SS

How can we stain?

Physical staining

Chemical staining

Monochrome stain Dichrome stain

How many colours do we need

MetachromasiaHow many colours do we need

T

Acid (-)S

Base (+)T

Base (+)S

Acid (-)T

Acidophilic

Basophilic

Do we understand basophilic and acidophilic?

Do we understand basophilic and acidophilic?

T

Acid (-)S

Base (+)P(NH)

Base (+)S

Acid (-)DNA

Acidophilic

Basophilic

2

Do we understand basophilic and acidophilic?

Acid (-)P(-cooh)

Base (+)S

Do we understand basophilic and acidophilic?

Do we understand basophilic and acidophilic?

Acidophilic or basophilic

Do we understand basophilic and acidophilic?

Acidophilic or basophilic

Do we understand basophilic and acidophilic?

Acidophilic and basophilic

Do we understand basophilic and acidophilic?

Acidophilic or basophilic

Do we understand basophilic and acidophilic?

Dark or pale

Does dark mean blue?

Dark or pale

Does dark mean blue?

Dark or pale

Does dark mean blue?

Dark or pale

Does dark mean blue?

Dark or pale

Does dark mean blue?

Dark or pale

Does dark mean blue?

Dark or pale

Does dark mean blue?

Acidophilic and basophilic

Can we identify tissue types with naked eye?

Can we identify tissue types with naked eye?

Can we identify tissue types with naked eye?

Inadequate staining

What problems do we face in staining?

Clearing by xylene

Uneven staining

What problems do we face in staining?

Proper timemaintaining and dehydration

Overstaining with hematoxylin

What problems do we face in staining?

AcidAlcohol

Understaining with eosin

What problems do we face in staining?

Addingacetic acid

Overstaining with eosin

What problems do we face in staining?

Washingwithaldehyde tap waterOr 95% alcohol

Opacity

What problems do we face in staining?

Completedehydration

Presence of black component in section

What problems do we face in staining?

Filteringstain

Is routine staining enough?

Collagen Fiber

H and E stain

Mallory stainMasson trichome

Is routine staining enough?

Silver StainH and E stain

Reticular fiber

Is routine staining enough?

H and E stain

Elastic fibre

Van gieson stain Weigert’s Resorcin fuchsin

Is routine staining enough?

Goblet cell

H and E

Alcian bluePAS stain

Is routine staining enough?

Glomerular basement membrane

PAS stainH and E stain

Is routine staining enough?

Glycocalyx membrane of villus

H and EPAS stain

Is routine staining enough?

H and E Mallory Azan

Islets of Langerhans

Is routine staining enough?

Congo red

Amyloid

H and E

Is routine staining enough?

Feulgen stains

DNA

H and E

Is routine staining enough?

Fat stain (osmium tetraoxide)Fat stain (Oil red O stain)

Fat

H and E

Is routine staining enough?

So, what did we discuss today?

Thank You

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