This document outlines the histotechnique process which tissues undergo before microscopic examination. Key steps include: fixation to preserve tissue structure; processing involving dehydration, clearing, and impregnation to allow sectioning; embedding tissues in paraffin blocks for microtomy; sectioning samples and staining, typically with hematoxylin and eosin, for visualization under the microscope. Finally, samples are mounted on slides and labeled for storage and pathological examination.
This is a presentation covering all techniques in histopathology. Comprehensive coverage of all related aspects.. Useful for postgraduate Pathology students and practitioners.
This is a presentation covering all techniques in histopathology. Comprehensive coverage of all related aspects.. Useful for postgraduate Pathology students and practitioners.
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.
Microtomy, or the preparation of tissue slides, is the foremost technique used in histological studies. This presentation is a brief overview of the technique and the steps involved.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
2. Histopathology
It is a branch of pathology which deals with
the study of disease in a tissue section.
The tissue undergoes a series of steps before it
reaches the examiners desk.
3. PROTOCOLS FOLLOWED IN
HISTOTECHNIQUES :
Receipt and identification
Labelling of the sample with a UNIQUE lab identification number
Fixation
Dehydration
Clearing
Impregnation
Embedding
Section cutting
Staining
Mounting
4. Type of material obtained in laboratory
The human tissue comes from the surgery and the
autopsy room.
From surgery two types of tissue are obtained.
1. A small piece of lesions or tumor which is sent for
diagnosis before final removal of the lesion or the
tumor (Incisional biopsy).
2. If the whole of the tumor or lesion is sent for
examination and diagnosis by the pathologist, it is
called excisional biopsy.
Tissues from the autopsy are sent for the study of
disease and its course.
6. As soon as the specimen is received in the
laboratory
Check label (name, age, Hospital Registration No. and
the nature of tissue to be examined, clinical diagnosis
and the requisition form is also duly filled.)
Check if the specimen is in proper fixative.
Check if the financial matters have been taken
care off.
Make the entries in biopsy register and give the
specimen a pathology number which is also
known as an accession number.
7. FIXATION
It is a complex series of chemical events which
brings about changes in the various chemical
constituents of cell like hardening, however the cell
morphology and structural detail is preserved.
Unless a tissue is fixed soon after the removal from
the body it will undergo degenerative changes due
to autolysis and putrefaction hence, tissue
architecture will be lost.
8. Principle of fixation- The fixative brings about
crosslinking of proteins which produces
denaturation or coagulation of proteins; so that
it maintains everything in their in vivo relation.
9. Aims & effects of fixation
The preservation of cells & tissue constituents in
as life-like manner as possible.
To check against bacterial contamination.
Hardening : the hardening effect of fixatives allows
easy manipulation of soft tissue like brain,
intestines etc.
Solidification: solidify colloid material.
10. Properties of fixatives
Coagulation and precipitation (as described above).
Penetrate rapidly.
Solubility of fixatives - All fixatives should be
soluble in a suitable solvent, preferably in water so
that adequate concentrations can be prepared.
Concentration - It is important that the
concentration of fixative is isotonic.(Osmosis)
Reaction - Most fixatives are acidic. It may help in
fixation but can affect staining so has to be
neutralized.
11. NO fixative will penetrate a piece of tissue thicker
than 1 cm hence while dealing with thicker specimen
following methods are recommended.
SOLID ORGAN :- cut slices as necessary ,but not
thicker than 5 mm.
HOLLOW ORGAN :- either open or pack lightly
with wool soaked in fixative
LARGE ORGAN :- It requires dissection, inject
fixative along the vessels or bronchi as in case of
lung so that it reaches all parts of the organ.
12. Common fixative used for fixation of tissue
10% neutral formalin
Cheap, easily available, penetrates rapidly
Does not over harden the tissue when kept indefinitely.
Time required for fixation 24-48 hrs.
Disadvantage : formalin pigment is formed in areas of
hemorrhage.
The amount of formalin required for fixation of tissues
10 –20 times the volume of the specimen.
13. Mechanism of action :
Act by forming cross link between amino acids of the
proteins thereby making them insoluble.
Factors affecting fixation:
Size & thickness of tissue.
Organs containing large amount of blood fix slowly.
Fatty & lipomatous tissues fix slowly.
Time required for fixation.
At room temperature - 12 hours, For small biopsies - 4-6
hours. At 65°C fixation occurs in - 2 hours
16. Decalcification
Bone is a hard tissue so it is difficult to cut by
microtome it hence needs to be softened before
sectioning. The process of softening is done by
removing the calcium from the bone. This is called
Decalcification.
Decalcifying Agents : Two major types are :
1) Acids
Strong acids :Nitric acid (5-10 % -commonest),
HCL
Weak acids : Formic acid, acetic acid, picric acid
2) Chelating Agents : EDTA
23. THE DIFFERENT STEPS IN TISSUE
PROCESSING
Fixation
Dehydration : stagnant water is a source of bacterial
contamination & tissue destruction. Water in cells is
removed with dehydrating agent . In ascending
grades of alcohol.
24. Clearing : removing alcohol on one side & replacing it
with paraffin on the other side. Xylene is used due to its
fairly rapid action. Other egs. Chloroform, Benzene, Carbon
tetrachloride, Cedar-wood oil.
Xylene:
clearing
Xylene
25. Impregnation : The empty spaces in the tissue ,after
removal of water by dehydration is filled with
paraffin wax. This is known as impregnation. Done
by immersing the tissue in two changes of molten
wax. Adv: to make the tissue firm in consistency for
cutting.
Done in an oven heated to 52-54 C
Paraplast (purified paraffin + plastic ploymers)
26. In order to prepare a tissue for embedding, it
must be infiltrated with paraffin. Because water
and paraffin are not miscible, the specimens
must be gradually dehydrated to achieve
replacement of water with alcohol before the
clearing agent is introduced.
Once successfully dehydrated, a clearing agent
that is miscible with alcohol and paraffin (i.e.
xylene ) is infiltrated through the tissue. Finally,
the paraffin is introduced and completes the
tissue for embedding.
27. Embedding : The impregnated tissue is put in a
mould & molten wax is added so as to make a wax
block to hold the tissue for cutting.
Wax with melting point of 54-56 C is used.
Once the tissue has been processed it is ready to be
orientated into a paraffin block and subsequently
sectioned.
Orientation during embedding is crucial for the
representation of proper morphology.
(structures in skin, small gastrointestinal biopsies, and
vas deferens are among those for which orientation is
especially critical.)
29. L-mould (Leuckhart’s embedding
mould)
Two ‘L’ shaped pieces of heavy brass metal can be
adjusted to make various size moulds.
L-moulds are kept on a glass. Tissue is placed on the
bottom & then the space is filled with molten wax.
The wax is cooled & solidified, L-moulds are
removed.
The brick like square with embedded tissue is
called paraffin block.
32. Tissue –Tek-system
Two parts
stainless steel base mould & plastic mould.
Tissue is placed in steel mould & filled with molten
wax.
Plastic mould is placed on top of steel mould & again
filled with molten wax.
Once the wax is cooled & solidified, steel mould is
taken out.
Plastic mould remains & act as a holder for microtome
for sectioning.
33. Microtome
A microtome (from the Greek mikros, meaning
“small”, and temnein, meaning “to cut”) is a tool used
to cut extremely thin slices of material, known as
sections.
TYPES OF MICROTOME
I. Sledge Microtome (Typical cut thickness achievable is
between 10 and 60 micron)
II. Rotary Microtome
III.Auto-cut Microtome
37. Staining
H & E staining( Hematoxylin & eosin staining)
Mount in DPX.
38.
39. Cryostat
Microtome is housed within an insulated,
thermostatically controlled refrigerated cabinet.
Few drops of gel is put on chuck . This freezes to
make an ice block, on the tip of which required
tissue is put.
Temp. maintained around -20 degree C.
40.
41. Frozen section
Rapid diagnosis of tumors
Demonstration of fat by special stains
Quick Demonstration of amyloid
Localization of immunoglobulins
Intraoperative guide to surgeon as to surgical
margins are free of tumor
42. Mountant
The final stage in the preparation of tissues for
microscopy is mounting.
DPX (DISTRENE, PLASTICISER,
XYLENE)
One of the most commonly used mountants, DPX is a
colourless, neutral medium in which most standard stains
are well preserved. It is prepared by dissolving the
common plastic, polystyrene, in a suitable hydrocarbon
solvent (usually xylene).
43. Labelling and storage of slides
Slide mounted sections are identified during
preparation by inscribing the slide with the tissue
accession number or suitable code using a diamond
marker or pencil (frosted slides)
slides are usually stored, standing on their short
side, in metal or plastic drawers. Labels need to be
fixed to the opposite end and orientated so the
details can be read with the slide in this upright
position.