The frozen section procedure allows for rapid microscopic analysis of tissue specimens. Tissue is frozen to preserve its structure and cellular components. This allows thin sections of the tissue to be cut without chemical processing. Frozen sections are used for rapid diagnosis during surgery and research applications where chemical fixation could damage antigens or enzymes. The cryostat machine houses a microtome that can precisely cut thin frozen sections at controlled temperatures for examination.
technique of preparing imprint smear# comparision with frozen sections# application and its role in thyroid ,paathyroid,breast,skin,head and neck and mucinous tumors# advantages and limitations
This presentation deals tissue processing in histopathology, the detailed of presentation given blow:
Histology, study the organization of tissues at all levels, from the whole organ down to the molecular components of cells that are found in most multicellular plants and animals.
Animal tissues are classified as epithelium, with closely spaced cells and very little intercellular space; connective tissue, with large amounts of intercellular material; muscle, specialized for contraction; and nerve, specialized for conduction of electrical impulses. Blood is also sometimes considered a separate tissue type.
Plants are composed of relatively undifferentiated tissue known as meristematic tissue; storage tissue or parenchyma; vascular tissue; photosynthetic tissue or chlorenchyma and support tissue or sclerenchyma and collenchyma.
The Indian Dental Academy is the Leader in continuing dental education , training dentists in all aspects of dentistry and
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technique of preparing imprint smear# comparision with frozen sections# application and its role in thyroid ,paathyroid,breast,skin,head and neck and mucinous tumors# advantages and limitations
This presentation deals tissue processing in histopathology, the detailed of presentation given blow:
Histology, study the organization of tissues at all levels, from the whole organ down to the molecular components of cells that are found in most multicellular plants and animals.
Animal tissues are classified as epithelium, with closely spaced cells and very little intercellular space; connective tissue, with large amounts of intercellular material; muscle, specialized for contraction; and nerve, specialized for conduction of electrical impulses. Blood is also sometimes considered a separate tissue type.
Plants are composed of relatively undifferentiated tissue known as meristematic tissue; storage tissue or parenchyma; vascular tissue; photosynthetic tissue or chlorenchyma and support tissue or sclerenchyma and collenchyma.
The Indian Dental Academy is the Leader in continuing dental education , training dentists in all aspects of dentistry and
offering a wide range of dental certified courses in different formats.for more details please visit
www.indiandentalacademy.com
Cryosectioning
Frozen section in surgical/anatomic pathology
General notion about the procedure of frozen section
Indication and limitations of the procedure in anatomic/surgical pathology
Presented at ISBER 2015, Dr. Alex Esmon, Senior Product Manager at Thermo Fisher Scientific explains how to manage the cold chain to minimize pre-analytical variability. In his presentation, he discusses three scenarios: the importance of maintaining sample conditions, concerns during inventorying, and sample transport (including unit transfer and content transfer). To read more about biobanking best practices, check out our Inside Biobanking blog here: http://acceleratingscience.com/biobanking/
Histopathology is examination of tissues for presence or absence of changes in their structure due to disease processes. We go through various steps in the process of converting gross sample to microscopic slides.
Ovarian tumors Lecture notes for MBBS.pptxSizan Thapa
Introduction to ovarian tumors, Epidemiology, Classification of ovarian tumor, Pathogenesis of epithelial ovarian tumors, Serous tumors of the ovaries, definition, pathogenesis, gross and microscopic pathology, Mucinous tumors of ovaries, definition, pathogenesis, gross and microscopic pathology, Teratoma of the ovaries,definition, pathogenesis, gross and microscopic pathology, Dysgerminma,definition, pathogenesis, gross and microscopic pathology
Hydrocele and tumors of testis Introduction.pptxSizan Thapa
Presentation for MBBS student. Short notes on Hydrocele, and Introduction of Testicular tumors, Etiopathogenesis, including the WHO classification of Testicular tumors, 5th Edition, 2022.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
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Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
2. ● The frozen section procedure is a pathological
laboratory procedure to perform rapid
microscopic analysis of a specimen.
● The technical name is CRYO-SECTION.
● It is a method that produces the sections
without dehydrating, clearing agents and
embedding media as well.
3. ● History
– Resulted from need for rapid intraoperative
Diagnosis.
– Mayo clinic chief of pathology Louis B Wilson
pioneered the frozen section in 1905.
4. ● Uses of Frozen section in histology laboratory
– Rapid production of sections for intraoperative
diagnosis.
– Diagnostic and research of enzyme histochemistry.
Enzymes are labile
– Immunofluorescent methods
– Immunohistochemistry methods: heat and fixation
may inactivate or destroy antigens.
– Diagnostic and research non-enzyme
histochemistry – eg lipids and carbohydrates.
– Silver methods used particularly in neuropathology.
5. ● Indications of frozen sections.
– Urgent diagnosis during surgery
– Determine the involvement of resection margins in
malignancy. Eg Basal cell carcinoma
– Identifying ganglion cells in Hirschprung disease.
– Enzyme histochemistry (ATPase, NADPH)
– Non-enzyme histochemistry (lipid, carbohydrate)
– For silver and gold impregnation methods
– Identifying type of tissue eg parathyroid.
6. ● Theoretical considerations
– Principle of frozen sections
● When the tissue is frozen, the water in the tissues turns
to ice. The tissue becomes firm in this state.
● The ice acts as the embedding media.
● The consistency of the frozen block can be altered by
altering the temperature of the tissue. Decreasing the
temperature produces harder block while increasing
temperature softens the block.
● Majority of non-fatty unfixed tissue section well at -25 deg
C.
● Sectioning of fixed tissue requires a block at -10 deg C.
7. ● Fixation
– Frozen section requires unfixed tissues.
– Post fixation – for enzyme histochemistry
(controlled conditions)
8. ● Freezing Techniques
– Liquid nitrogen ( -190 deg C)
– Isopentane cooled by liquid nitrogen ( - 150 deg C)
– Dry ice ( -70 deg C)
– Carbon dioxide ( -70 deg C)
– Aerosol sprays ( - 50 deg C)
9.
10. ● Methods for suitable freezing
– Cryostat
● Microtome is inside the chamber
● Microtome is temperature controlled
– Freezing microtome
● Tissue fixed separately
● Microtome is not under temperature control.
11. Cryostat
● The cryostat is a refrigerated cabinet in which a
specialty microtome is housed.
● All the controls to the microtome is operated
outside the cabinet.
● Features
– Electronic temperature control
– Electronically controlled advance and retraction of
the block.
– Specimen orientation facility
12.
13. – Digital visualization of chuck and cabinet
temperature.
– Mechanical cutting speed control and section
thickness
– Automatic defrost mechanism
– Automatic de-contamination and sterilization.
15. ● The best quality frozen sections are produced
from fresh unfixed sections that has been
rapidly frozen.
● Cryostat freezing may produce freezing artifact
because of slower freezing than other
techniques.
● The water in the tissues form ice crystal when
the tissue freezes.
16. ● The size and quality of the crystals is proportional to
the speed at which the tissue is frozen.
● After cutting, the sections are placed on a room
temp slide → thawing of the tissue → produces
freezing artifact appearing as holes in the tissue
when viewed microscopically.
● Method of choice of freezing.
– Isopentane and
– liquid nitrogen (problem : produces vapor bubbles around
the tissue → acts as insulator → prevents rapid cooling.
→ freeze artifact.
17. ● To localize hydrolytic enzymes eg acid and
alkaline phosphatase, and other antigens, the
tissue is fixed prior to freezing and sectioning.
● Method
– Tissue must be fresh
– Place in formal calcium at 4 deg C for 18 hours for
fixation.
– Rinse in running water or Distilled water(if the tissue
is fragile)
18. – Blot dry
– Place tissue in gum sucrose solution for 18 hrs at 4
deg C
– Blot dry
– Freeze tissue onto block holder
19. ● Cryostat sectioning
– Cabinet temperature
● Should be suitable for tissue type and type of preparation
to be cut
● Most unfixed material : -15 deg C to -23 deg C
● Tissue containing large amount of water or fixed tissues
section best at warmer temperature. ( -7 to -12 deg C)
● Tissue containing fat require a colder temperature.
● Shattered sections – block is too cold.
20. ● Microtome
– In case of cutting problems – defrost microtome,
and oil as recommended.
● Blade/knife
– Disposable blades have replaced stainless steel
knives.
– Type of tissue and procedure to be performed
dictate the use of knives.
– Sharpening techniques should be mentioned in
procedure manual.
– Sharp edge is essential for proper section.
21. ● Disposable blades have perfect edge and
instantly available.
● They have an advantage of sharpness, and the
blades are rapidly cooled due to their size.
● Hard or dense tissue may be troublesome for
disposable blades.
22. ● Anti-roll plate
– Piece of equipment attached to the front of
microtome.
– Prevents the upward curling of frozen section
during sectioning.
– Made of plexiglass or hard plastic.
– Aligned parallel to the blade and slightly above it.
23. ● Sectioning technique
– Speed, tissue type, and temperature of the block
and the cabinet is vital in sectioning.
– After cutting, the cut section will rest on the blade
holder.
– Room temperature slide is held above the section.
Electrostatic attraction causes the tissue to adhere
to the slide.
– If staining technique to be used is lengthy, positively
charged slides should be used.
24. ● Coatings
– Gelatin – formladehyde mixture
● Gelatin 1% (5ml)
● Formaldehyde 2% (5ml)
● Coat slides, allow to dry at 37 deg C for 1 hr or overnight
before picking up sections.
– Poly – L – lysin coating (0.01% PLL aqueous)
● Wash slides in detergent – 30 min
● Wash slides in running tap water – 30 min
● Rinse slides in DW two times – 5 min each
25. ● Wash slides in 95% alcohol twice – 5 min each
● Air dry slides – 10 min
● Smear 20 micro liter of PLL over each slide
● Air dry and store dust free
This technique is used as section adhesive in
immunohistochemistry.
26. ● RAPID BIOPSY FOR INTRAOPERATIVE
DIAGNOSIS
– Frozen section is valuable for rapid diagnosis during
surgery.
– Selected tissue is frozen using one of the freezing
techniques.
– Slide is immediately immersed in cold acetone or 95%
alcohol.
– Sections are immediately stained by rapid H&E,
methylene blue, or polychrome stain.
27. ● Ultracryotomy
– Used in research laboratory.
– Rapid freezing of fixed or unfixed tissue using
isopentane or liquid nitrogen.
– Sections cut at 50 -150 nm.
28. ● Freeze drying and freeze substitution.
– Freeze drying is a technique of rapid freezing
(quenching) at -160 deg C and subsequent removal
of water molecules by sublimation in a vacuum at a
higher temperature (-40 deg C).
– The blocks are raised to room temperature and
fixed by vapor.
– Advantages – this technique minimizes :
● Loss of soluble substances.
29. ● Displacement of cell constituents.
● Chemical alteration of reactive groups
● Denaturation of proteins
● Destruction or inactivation of enzymes.