SlideShare una empresa de Scribd logo
1 de 48
Descargar para leer sin conexión
Mr.Mahmoud Ibrahim




TEM is a microscopy technique
whereby a beam of electrons is
transmitted through an ultrathin
specimen, interacting with the
specimen as it passes through it.
Introduction

An image is formed from the electrons
transmitted through the specimen,
magnified and focused by an objective
lens and appears on an imaging screen.

The standard approach is to immerse
the specimen in fixative pre-cooled to
4oc immediately after collection.
Specimen handling

Once in the fixative the specimen is
selected using scalpel, then transferred
to glass vial.
The final specimen size is usually
1mm3.

Post mortem changes and fixation
artefact reduced with small samples.

The standard fixation protocol involves
primary fixation with glutaraldehyde at
40c followed by secondary fixation in
osmium tetroxide.
Fixation

Glutaraldehyde is effective at a
concentration 1.5-4%, while osmium
tetroxide optimum concentration 1 or
2%.
Fixation concentration

Optimum temperature 4oc.
Fixation at room temperature improve
the penetration rate, reduce the time but
increases the risk of autolytic change.
Fixation temperature

For 0.5-1mm3 tissue specimen in
aldehyde fixatives 4-6 hrs.
Secondary fixation in osmium tetroxide
required 60-90 minutes.
Fixation duration

Optimum pH ranged between 7.2-7.6.
Inadequate buffering can lead to
significant loss of cellular components,
with subsequent shrinkage or swelling.
Fixation pH

Generally is not a major requirement.
If necessary 300-330 mOsm.
Osmolality equivalent to that of plasma
or slightly hypertonic.
Fixation osmolality

Rinsing the tissue in buffer after post
fixation.
Done to remove surplus fixative and
provide an opportunity for tissue storage
if necessary.
Wash buffer

Dehydration usually carried out in
ethanol with the combination of
propylene oxide to facilitate resin
infiltration.
Dehydration

The dehydrating times should be
adjusted to size and kind of tissue.

Optimum impregnation as
follows:
First place tissue in a solution
composed of equal volume from
propylene oxide and resin.
Impregnation

Second transfer tissue to 25:75
propylene oxide and resin.
Finally put tissue in pure resin.
An hour in each step is adequate.

Periodic agitation is
recommended in each step.
Incomplete impregnation causes
major sectioning difficulties.

Tissue samples are placed in an
appropriate mold (capsules
made from polyethylene glycol)
filled with resin and allowed to
polymerized using heat.
Embedding

A paper strip bearing the tissue
identification code written in
pencil should be used.
Flat embedding molds made from
silicone can be used.

The most common embedding media
for E.M are:
1- Epoxy resins.
2- Acrylic resins.
Embedding media

Characterized by:
1- Contains oxygen and 2 carbon
atoms (epoxide), cross linking
between them creates a three
dimensional polymer of great
mechanical strength.
Epoxy resins

2- With little shrinkage (less
than 2%).
3- Permanent.
4- Preserve tissue ultra structure.

5- Stable in E.M.
6- Easy section.
7- Available.

Epoxy resin composed of:
1- Monomeric resin.
2- Hardener.
3- Accelerator.
4- Plasticizer.

Hardness, flexibility and
polymerization times can be
manipulated by varying the
amount of individual
components

The prepared resin is best
delivered through a non-
reactive plastic syringe or
pipette.

The widely used epoxy resins
are:
1- Araldite.
2- Epon.
3- Spurr,s resin.

The exposure to epoxy resins
causes allergic contact dermatitis.
May be carcinogenic.
Toxic.
Irritants.

Derive from methacrylic acid
and acrylic acid.
Used to infiltration and
embedding as epoxy resins.
Acrylic resins (methacrylates)

Polymerization obtained by:
light, heat or chemical
accelerator at room temperature

The most common acrylic resins
are:
LR white, LR gold and the
Lowicryl series (K4M, K11M,
HM20 and HM23).

Manual tissue processing is
preferably for E.M.
Cell suspensions can be
demonstrated by E.M.
Tissue processing

Glass or diamond knives used.
Higher angle knives up to 550 for
hard tissue while 350 for soft
tissue.
Ultra microtomy

D.W or deionized water used.
10-15% of ethanol or acetone can
also be used but not with a
diamond knife.
Floatation

The floatation fluid should be
attached to the knife

Can be achieved manually or by
using ultra microtome.
Trimming

Allow samples to be screened
for specific features and to
select areas for thin sectioning.
Carried out by using glass not
diamond knife.
Semi-thin section

Semi-thin sections usually
measured between 0.5-1
micrometer.

Usual section thickness 0.1-
0.005 micrometers.
Optimum section thickness
measures 80 nm.
Ultra-thin sectioning

To collect sections immerse the
grid in the floatation fluid, then
placed the ribbon over the grid.

After sections collection, the
grids transferred by using forceps
to filter paper on a Petri dish, and
allow to dry completely before
staining.

Achieved by introducing heavy
metal atoms which deposit on
the tissue components.
Staining

Tissues are stained at several
points during preparation:
1- During secondary fixation.

2- When uranyl acetate is used
during the post fixation wash.
3- By staining the sections with
lead and uranium salts.

Más contenido relacionado

La actualidad más candente

La actualidad más candente (20)

Fixation
Fixation Fixation
Fixation
 
Fixation of tissues
Fixation of tissuesFixation of tissues
Fixation of tissues
 
Transmission electron microscope
Transmission electron microscopeTransmission electron microscope
Transmission electron microscope
 
Principles, Methods and Types of Fixatives
Principles, Methods and Types of FixativesPrinciples, Methods and Types of Fixatives
Principles, Methods and Types of Fixatives
 
Microtomy
MicrotomyMicrotomy
Microtomy
 
Autoradiography
AutoradiographyAutoradiography
Autoradiography
 
Polarizing Light Microscopy
Polarizing Light MicroscopyPolarizing Light Microscopy
Polarizing Light Microscopy
 
Hematoxylin and Eosin Staining
Hematoxylin and Eosin StainingHematoxylin and Eosin Staining
Hematoxylin and Eosin Staining
 
Mictome
MictomeMictome
Mictome
 
Fluorescence microscope
Fluorescence microscopeFluorescence microscope
Fluorescence microscope
 
Principles of Staining
Principles of StainingPrinciples of Staining
Principles of Staining
 
Light microscope
Light microscopeLight microscope
Light microscope
 
Fluoresence microscope
Fluoresence microscopeFluoresence microscope
Fluoresence microscope
 
Tissue preparation process, embedding and block preparation in research area
Tissue preparation process, embedding and block preparation in research areaTissue preparation process, embedding and block preparation in research area
Tissue preparation process, embedding and block preparation in research area
 
Electron microscope ppt
Electron microscope pptElectron microscope ppt
Electron microscope ppt
 
Electron microscope, principle and application
Electron microscope, principle and applicationElectron microscope, principle and application
Electron microscope, principle and application
 
Fluorescence Microscopy
Fluorescence MicroscopyFluorescence Microscopy
Fluorescence Microscopy
 
Histological tools
Histological toolsHistological tools
Histological tools
 
Hematoxylin and Eosin staining for UGs
Hematoxylin and Eosin staining for UGsHematoxylin and Eosin staining for UGs
Hematoxylin and Eosin staining for UGs
 
Autoradiography
AutoradiographyAutoradiography
Autoradiography
 

Similar a Electron microscope

Transmission electron microscope
Transmission electron microscopeTransmission electron microscope
Transmission electron microscopeSizan Thapa
 
Specimen Preperation for Electron Microscope .pptx
Specimen Preperation for Electron Microscope .pptxSpecimen Preperation for Electron Microscope .pptx
Specimen Preperation for Electron Microscope .pptxSreedevi694198
 
PREPARATION OF HISTOLOGICAL SPECIMENS.pptx
PREPARATION OF HISTOLOGICAL SPECIMENS.pptxPREPARATION OF HISTOLOGICAL SPECIMENS.pptx
PREPARATION OF HISTOLOGICAL SPECIMENS.pptxAnthonyMatu1
 
Tissue processing 2012
Tissue processing 2012Tissue processing 2012
Tissue processing 2012Dhiraj Shukla
 
Support Films for Electron Microscopy Grids
Support Films for Electron Microscopy  GridsSupport Films for Electron Microscopy  Grids
Support Films for Electron Microscopy GridsDr. Fatma Kassem
 
Transmission Electron Microscopy
Transmission Electron MicroscopyTransmission Electron Microscopy
Transmission Electron MicroscopyMaha Hammady
 
5 lecture preparation histological specimens.pdf
5  lecture preparation histological specimens.pdf5  lecture preparation histological specimens.pdf
5 lecture preparation histological specimens.pdfAroo9
 
Histokinetic by dr narmada
Histokinetic by  dr narmadaHistokinetic by  dr narmada
Histokinetic by dr narmadaNarmada Tiwari
 
Technique of Histopathology
Technique of HistopathologyTechnique of Histopathology
Technique of Histopathologysandeep singh
 
Technique of Histopathology
Technique of HistopathologyTechnique of Histopathology
Technique of Histopathologysandeep singh
 
Histological techniques
Histological techniquesHistological techniques
Histological techniquesRUPALIMUNDE
 
Fyp slide polymer solar cells
Fyp slide polymer solar cells Fyp slide polymer solar cells
Fyp slide polymer solar cells Faizzwan Fazil
 
DENTURE BASE RESIN
DENTURE BASE RESINDENTURE BASE RESIN
DENTURE BASE RESINdrfarhana4
 

Similar a Electron microscope (20)

TISSUE PROCESSING SEMINAR
TISSUE PROCESSING SEMINARTISSUE PROCESSING SEMINAR
TISSUE PROCESSING SEMINAR
 
Transmission electron microscope
Transmission electron microscopeTransmission electron microscope
Transmission electron microscope
 
Specimen Preperation for Electron Microscope .pptx
Specimen Preperation for Electron Microscope .pptxSpecimen Preperation for Electron Microscope .pptx
Specimen Preperation for Electron Microscope .pptx
 
PREPARATION OF HISTOLOGICAL SPECIMENS.pptx
PREPARATION OF HISTOLOGICAL SPECIMENS.pptxPREPARATION OF HISTOLOGICAL SPECIMENS.pptx
PREPARATION OF HISTOLOGICAL SPECIMENS.pptx
 
Tissue processing 2012
Tissue processing 2012Tissue processing 2012
Tissue processing 2012
 
Support Films for Electron Microscopy Grids
Support Films for Electron Microscopy  GridsSupport Films for Electron Microscopy  Grids
Support Films for Electron Microscopy Grids
 
Transmission Electron Microscopy
Transmission Electron MicroscopyTransmission Electron Microscopy
Transmission Electron Microscopy
 
Histotechniques
HistotechniquesHistotechniques
Histotechniques
 
5 lecture preparation histological specimens.pdf
5  lecture preparation histological specimens.pdf5  lecture preparation histological specimens.pdf
5 lecture preparation histological specimens.pdf
 
Histokinetic by dr narmada
Histokinetic by  dr narmadaHistokinetic by  dr narmada
Histokinetic by dr narmada
 
Elastomeric Fiber
Elastomeric Fiber Elastomeric Fiber
Elastomeric Fiber
 
Technique of Histopathology
Technique of HistopathologyTechnique of Histopathology
Technique of Histopathology
 
Technique of Histopathology
Technique of HistopathologyTechnique of Histopathology
Technique of Histopathology
 
Histological techniques
Histological techniquesHistological techniques
Histological techniques
 
Fyp slide polymer solar cells
Fyp slide polymer solar cells Fyp slide polymer solar cells
Fyp slide polymer solar cells
 
Plastination
Plastination Plastination
Plastination
 
Pod
PodPod
Pod
 
Acrulic synthtics fiber
Acrulic synthtics fiberAcrulic synthtics fiber
Acrulic synthtics fiber
 
DENTURE BASE RESIN
DENTURE BASE RESINDENTURE BASE RESIN
DENTURE BASE RESIN
 
Histopathology Procedures
Histopathology ProceduresHistopathology Procedures
Histopathology Procedures
 

Más de MAHMOUD IBRAHIM (20)

Carbohydrates
CarbohydratesCarbohydrates
Carbohydrates
 
2 workflow
2 workflow2 workflow
2 workflow
 
1 introduction
1 introduction1 introduction
1 introduction
 
biochemical hazard signs
 biochemical hazard signs  biochemical hazard signs
biochemical hazard signs
 
lab safety
 lab safety  lab safety
lab safety
 
Healing and repair
Healing and repairHealing and repair
Healing and repair
 
Frozen section and cryostat
Frozen section and cryostatFrozen section and cryostat
Frozen section and cryostat
 
Tissue microarray
Tissue microarrayTissue microarray
Tissue microarray
 
Museum
MuseumMuseum
Museum
 
Chronic inflammation
Chronic inflammationChronic inflammation
Chronic inflammation
 
Inflammation
InflammationInflammation
Inflammation
 
Mounting media
Mounting mediaMounting media
Mounting media
 
Introduction of pathology
Introduction of pathologyIntroduction of pathology
Introduction of pathology
 
Lipid
LipidLipid
Lipid
 
Hematoxylin and Eosin
Hematoxylin and EosinHematoxylin and Eosin
Hematoxylin and Eosin
 
Theory of stain
Theory of stainTheory of stain
Theory of stain
 
Connective Tissue
Connective TissueConnective Tissue
Connective Tissue
 
Demonstration of protein and nucleic acids
Demonstration of protein and nucleic acidsDemonstration of protein and nucleic acids
Demonstration of protein and nucleic acids
 
Carbohydrate
CarbohydrateCarbohydrate
Carbohydrate
 
Connective tissues
Connective tissuesConnective tissues
Connective tissues
 

Último

AUDIENCE THEORY -- FANDOM -- JENKINS.pptx
AUDIENCE THEORY -- FANDOM -- JENKINS.pptxAUDIENCE THEORY -- FANDOM -- JENKINS.pptx
AUDIENCE THEORY -- FANDOM -- JENKINS.pptxiammrhaywood
 
Diploma in Nursing Admission Test Question Solution 2023.pdf
Diploma in Nursing Admission Test Question Solution 2023.pdfDiploma in Nursing Admission Test Question Solution 2023.pdf
Diploma in Nursing Admission Test Question Solution 2023.pdfMohonDas
 
Philosophy of Education and Educational Philosophy
Philosophy of Education  and Educational PhilosophyPhilosophy of Education  and Educational Philosophy
Philosophy of Education and Educational PhilosophyShuvankar Madhu
 
M-2- General Reactions of amino acids.pptx
M-2- General Reactions of amino acids.pptxM-2- General Reactions of amino acids.pptx
M-2- General Reactions of amino acids.pptxDr. Santhosh Kumar. N
 
How to Show Error_Warning Messages in Odoo 17
How to Show Error_Warning Messages in Odoo 17How to Show Error_Warning Messages in Odoo 17
How to Show Error_Warning Messages in Odoo 17Celine George
 
Patient Counselling. Definition of patient counseling; steps involved in pati...
Patient Counselling. Definition of patient counseling; steps involved in pati...Patient Counselling. Definition of patient counseling; steps involved in pati...
Patient Counselling. Definition of patient counseling; steps involved in pati...raviapr7
 
CAULIFLOWER BREEDING 1 Parmar pptx
CAULIFLOWER BREEDING 1 Parmar pptxCAULIFLOWER BREEDING 1 Parmar pptx
CAULIFLOWER BREEDING 1 Parmar pptxSaurabhParmar42
 
How to Solve Singleton Error in the Odoo 17
How to Solve Singleton Error in the  Odoo 17How to Solve Singleton Error in the  Odoo 17
How to Solve Singleton Error in the Odoo 17Celine George
 
DUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRA
DUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRADUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRA
DUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRATanmoy Mishra
 
2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx
2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx
2024.03.23 What do successful readers do - Sandy Millin for PARK.pptxSandy Millin
 
UKCGE Parental Leave Discussion March 2024
UKCGE Parental Leave Discussion March 2024UKCGE Parental Leave Discussion March 2024
UKCGE Parental Leave Discussion March 2024UKCGE
 
Practical Research 1: Lesson 8 Writing the Thesis Statement.pptx
Practical Research 1: Lesson 8 Writing the Thesis Statement.pptxPractical Research 1: Lesson 8 Writing the Thesis Statement.pptx
Practical Research 1: Lesson 8 Writing the Thesis Statement.pptxKatherine Villaluna
 
PISA-VET launch_El Iza Mohamedou_19 March 2024.pptx
PISA-VET launch_El Iza Mohamedou_19 March 2024.pptxPISA-VET launch_El Iza Mohamedou_19 March 2024.pptx
PISA-VET launch_El Iza Mohamedou_19 March 2024.pptxEduSkills OECD
 
Drug Information Services- DIC and Sources.
Drug Information Services- DIC and Sources.Drug Information Services- DIC and Sources.
Drug Information Services- DIC and Sources.raviapr7
 
How to Add Existing Field in One2Many Tree View in Odoo 17
How to Add Existing Field in One2Many Tree View in Odoo 17How to Add Existing Field in One2Many Tree View in Odoo 17
How to Add Existing Field in One2Many Tree View in Odoo 17Celine George
 
The Stolen Bacillus by Herbert George Wells
The Stolen Bacillus by Herbert George WellsThe Stolen Bacillus by Herbert George Wells
The Stolen Bacillus by Herbert George WellsEugene Lysak
 
Benefits & Challenges of Inclusive Education
Benefits & Challenges of Inclusive EducationBenefits & Challenges of Inclusive Education
Benefits & Challenges of Inclusive EducationMJDuyan
 
P4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdf
P4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdfP4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdf
P4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdfYu Kanazawa / Osaka University
 
Presentation on the Basics of Writing. Writing a Paragraph
Presentation on the Basics of Writing. Writing a ParagraphPresentation on the Basics of Writing. Writing a Paragraph
Presentation on the Basics of Writing. Writing a ParagraphNetziValdelomar1
 

Último (20)

AUDIENCE THEORY -- FANDOM -- JENKINS.pptx
AUDIENCE THEORY -- FANDOM -- JENKINS.pptxAUDIENCE THEORY -- FANDOM -- JENKINS.pptx
AUDIENCE THEORY -- FANDOM -- JENKINS.pptx
 
Diploma in Nursing Admission Test Question Solution 2023.pdf
Diploma in Nursing Admission Test Question Solution 2023.pdfDiploma in Nursing Admission Test Question Solution 2023.pdf
Diploma in Nursing Admission Test Question Solution 2023.pdf
 
Philosophy of Education and Educational Philosophy
Philosophy of Education  and Educational PhilosophyPhilosophy of Education  and Educational Philosophy
Philosophy of Education and Educational Philosophy
 
M-2- General Reactions of amino acids.pptx
M-2- General Reactions of amino acids.pptxM-2- General Reactions of amino acids.pptx
M-2- General Reactions of amino acids.pptx
 
How to Show Error_Warning Messages in Odoo 17
How to Show Error_Warning Messages in Odoo 17How to Show Error_Warning Messages in Odoo 17
How to Show Error_Warning Messages in Odoo 17
 
Patient Counselling. Definition of patient counseling; steps involved in pati...
Patient Counselling. Definition of patient counseling; steps involved in pati...Patient Counselling. Definition of patient counseling; steps involved in pati...
Patient Counselling. Definition of patient counseling; steps involved in pati...
 
CAULIFLOWER BREEDING 1 Parmar pptx
CAULIFLOWER BREEDING 1 Parmar pptxCAULIFLOWER BREEDING 1 Parmar pptx
CAULIFLOWER BREEDING 1 Parmar pptx
 
How to Solve Singleton Error in the Odoo 17
How to Solve Singleton Error in the  Odoo 17How to Solve Singleton Error in the  Odoo 17
How to Solve Singleton Error in the Odoo 17
 
DUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRA
DUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRADUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRA
DUST OF SNOW_BY ROBERT FROST_EDITED BY_ TANMOY MISHRA
 
2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx
2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx
2024.03.23 What do successful readers do - Sandy Millin for PARK.pptx
 
UKCGE Parental Leave Discussion March 2024
UKCGE Parental Leave Discussion March 2024UKCGE Parental Leave Discussion March 2024
UKCGE Parental Leave Discussion March 2024
 
Practical Research 1: Lesson 8 Writing the Thesis Statement.pptx
Practical Research 1: Lesson 8 Writing the Thesis Statement.pptxPractical Research 1: Lesson 8 Writing the Thesis Statement.pptx
Practical Research 1: Lesson 8 Writing the Thesis Statement.pptx
 
PISA-VET launch_El Iza Mohamedou_19 March 2024.pptx
PISA-VET launch_El Iza Mohamedou_19 March 2024.pptxPISA-VET launch_El Iza Mohamedou_19 March 2024.pptx
PISA-VET launch_El Iza Mohamedou_19 March 2024.pptx
 
Drug Information Services- DIC and Sources.
Drug Information Services- DIC and Sources.Drug Information Services- DIC and Sources.
Drug Information Services- DIC and Sources.
 
How to Add Existing Field in One2Many Tree View in Odoo 17
How to Add Existing Field in One2Many Tree View in Odoo 17How to Add Existing Field in One2Many Tree View in Odoo 17
How to Add Existing Field in One2Many Tree View in Odoo 17
 
The Stolen Bacillus by Herbert George Wells
The Stolen Bacillus by Herbert George WellsThe Stolen Bacillus by Herbert George Wells
The Stolen Bacillus by Herbert George Wells
 
Benefits & Challenges of Inclusive Education
Benefits & Challenges of Inclusive EducationBenefits & Challenges of Inclusive Education
Benefits & Challenges of Inclusive Education
 
Personal Resilience in Project Management 2 - TV Edit 1a.pdf
Personal Resilience in Project Management 2 - TV Edit 1a.pdfPersonal Resilience in Project Management 2 - TV Edit 1a.pdf
Personal Resilience in Project Management 2 - TV Edit 1a.pdf
 
P4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdf
P4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdfP4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdf
P4C x ELT = P4ELT: Its Theoretical Background (Kanazawa, 2024 March).pdf
 
Presentation on the Basics of Writing. Writing a Paragraph
Presentation on the Basics of Writing. Writing a ParagraphPresentation on the Basics of Writing. Writing a Paragraph
Presentation on the Basics of Writing. Writing a Paragraph
 

Electron microscope