Liquid based cytology (LBC) has been introduced to improve cervical screening. It uses a liquid preservative instead of smearing cells directly on a slide. This allows automated processing to disperse cells and transfer them evenly to a slide. LBC reduces sampling errors, improves cell preservation and slide quality compared to conventional smears. However, it requires specialized equipment and is more expensive. Laser scanning cytometry is a technique that can analyze individual cells from LBC samples. It provides quantitative measurements and complements flow cytometry by allowing analysis of adherent cells and solid tissues.
Cervical cytology was introduced by George
Papanicolaou into clinical practice in 1940. In 1945,
the Papanicolaou smear received the endorsement of
the American cancer society as an effective method
for the prevention of cervical cancer .
www.drvikramsaraswat.co.in
www.drsaraswatpathlabs.com
cytology of urine tract - this slide contains the specimen collection method, preparation of specimen, types of fixatives, other preparation techniques, urinary tract histology, normal urinary tract cytology,
Cervical cytology was introduced by George
Papanicolaou into clinical practice in 1940. In 1945,
the Papanicolaou smear received the endorsement of
the American cancer society as an effective method
for the prevention of cervical cancer .
www.drvikramsaraswat.co.in
www.drsaraswatpathlabs.com
cytology of urine tract - this slide contains the specimen collection method, preparation of specimen, types of fixatives, other preparation techniques, urinary tract histology, normal urinary tract cytology,
Urine analysis is an integral part of a clinical laboratory. automation techniques in urine biochemistry, their priniciplas and microscopy along with their advantages and disadvantages are outlined.
Liquid-based cytology is a method of preparing samples for examination in cytopathology. The sample is collected, normally by a small brush, in the same way as for a conventional smear test, but rather than the smear being transferred directly to a microscope slide, the sample is deposited into a small bottle of preservative liquid. At the laboratory the liquid is treated to remove other elements such as mucus before a layer of cells is placed on a slide. The technique allows more accurate results. The UK screening programmes changed their cervical screening method from the Pap test to liquid-based cytology in 2008.
Atlas on bethesda system for reporting cervical cytologyAshish Jawarkar
This is an atlas with more nearly 100 images, authentic taken from NCI web atlas. Useful to understand and report pap smears. The subject has been presented in a way which will help students reproduce in exams.
The technique of flow cytometry is used to evaluate cells for a number of functions, such as cell counting, phenotyping, cell cycle analysis, and viability.
Urine analysis is an integral part of a clinical laboratory. automation techniques in urine biochemistry, their priniciplas and microscopy along with their advantages and disadvantages are outlined.
Liquid-based cytology is a method of preparing samples for examination in cytopathology. The sample is collected, normally by a small brush, in the same way as for a conventional smear test, but rather than the smear being transferred directly to a microscope slide, the sample is deposited into a small bottle of preservative liquid. At the laboratory the liquid is treated to remove other elements such as mucus before a layer of cells is placed on a slide. The technique allows more accurate results. The UK screening programmes changed their cervical screening method from the Pap test to liquid-based cytology in 2008.
Atlas on bethesda system for reporting cervical cytologyAshish Jawarkar
This is an atlas with more nearly 100 images, authentic taken from NCI web atlas. Useful to understand and report pap smears. The subject has been presented in a way which will help students reproduce in exams.
The technique of flow cytometry is used to evaluate cells for a number of functions, such as cell counting, phenotyping, cell cycle analysis, and viability.
This presentation in mainly focused of understanding of automation and its utility in cytopathology. It will be very usefull for postgraduate in pathology, cytopathologist and cytotechnicians.
Fluorescence- Activated Cell Sorter is a powerful technique used in cell sorting, cell-cycle analysis etc.
The presentation gives a basic understanding of the principle of FACS, instrumentation, interpretation of results, applications, how to do cell-cycle analysis using FACS and various troubleshooting tips.
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
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
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.
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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!
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
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The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
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The Gram stain is a fundamental technique in microbiology used to classify bacteria based on their cell wall structure. It provides a quick and simple method to distinguish between Gram-positive and Gram-negative bacteria, which have different susceptibilities to antibiotics
2. Background
• The conventional pap smear
has been the most successful
screening test
• Screening every 3-5years has
resulted in a 70% reduction in
incidence
3. Why LBC (Liquid based cytology) has
been introduced ?
• Continuing improvement to the Cervical
screening Programme
• Limitations of conventional cytology
• Modernisation of the technique
• Future benefits
– Extra tests – HPV, chlamydia, Neiseria
Gonorrhoea.
4. Limitations Of Conventional Smear
(From UK studies)
False Negative Rate of up to 55%1
Sampling and interpretive errors
Ambiguous reports of 6.4%2
70% are truly negative
30% represent more severe abnormality
Inadequate specimens of 9.7% 2
1. Hutchinson et al., AJCP, Vol 101-2; 215-219, 2. DOH Statistical Bulletin 2000/2001
5. Sources Of False Negatives
• Sampling issues (70%)
– cells not collected on the sampling device
– cells collected, but not transferred to the slide
• Thicker and thinner areas
• Nuclear feathering artifact
• Interpretive issues (30%)
– abnormal cells present on slide but either not seen
or misinterpreted
• Blood/mucus
• Air drying artifact
6. What does 'Liquid Based Cytology'
mean?
• Literally it means
“cytology (the study of cells)
through a liquid medium.”
11. Specimen Collection
• PreservCyt Solution.
• Capped, labeled, and sent to the
laboratory equipped with a
ThinPrep 2000 Processor .
Composition
• Buffered methanol
• No active ingredient
Storage
• 15 to 30 C for 6 weeks
13. Thin Prep processor
(1)Cell dispersion
• Swirling the sampling device in
the preservation solution
• Strong enough to separate
debris and disperse mucus, but
gentle enough to have no
adverse effect on cell
appearance.
14. Thin Prep processor
(2) Cell Collection
• A gentle vacuum is created
within the ThinPrep Pap Test
Filter, which collects cells on the
exterior surface of the membrane.
• Cell collection is controlled by the
ThinPrep 2000 Processor’s
software that monitors the rate of
flow through the ThinPrep Pap
Test Filter.
15. Thin Prep processor
(3) Cell Transfer
• After the cells are collected on
the membrane, the ThinPrep
Pap Test Filter is inverted and
gently pressed against the
ThinPrep Microscope Slide.
• Natural attraction and slight
positive air pressure cause the
cells to adhere to the ThinPrep
Microscope Slide resulting in an
even distribution of cells in a
defined circular area.
33. Procedure
Decant fixative
Allow 3-6 drops
Collection of and blot
of suspension to
sample excessive
glass slide
fixative
Add 1-2 ml of
Specimen is
polymer solution Allow to dry
vortex mixed
to tube
Centrifuge at Stain with conv.
Vortex mix
800 g for 10 min Pap
35. What is the need?
• Our limited ability to undertake accurate,
quantitative measurement of cellular and
subcellular factors
• Established technologies in clinical pathology,
including conventional microscope-based
histopathology and histochemistry,
fluorescence microscopy, flow cytometry and
computer-based image cytometry, all have
limitations.
36. Introduction
• Imaging + cytometric analysis
• Not random, but event-based.
• It is closely related to conventional flow
cytometry, which also analyzes
individual cells that meet certain
characteristics (and is also event-
based). Both are therefore cytometric
techniques
37. Limitations of Flow cytometry
1. time-resolved events such as enzyme
kinetics cannot be analyzed.
2. Simultaneous study of Morphology of
the measured cell is not possible.
3. Cell analysis is at zero spatial
resolution.
4. The cell once measured cannot be re-
analyzed with another probe(s)
38. 5. Analysis of solid tissue requires cell or
nucleus isolation, a procedure that may
produce artifacts and loss of the
information on tissue architecture.
6.size samples such as fine needle
aspirates, spinal fluid, thus, are seldom
analyzed by FC.
7. The measured sample cannot be stored
for archival preservation.
39. Introduction
• 2 manufacturers:
– CompuCyte Corp. (Cambridge, MA)
– Olympus Optical Co. (Tokyo), offers many
advantages of flow cytometry but has no
limitations listed above.
• The analytical capabilities of LSC, therefore,
complement these of FC, and extend the use
of cytometry in many applications
45. Thresholding on flow cytometer
• Setting the threshold (or discrimination) on the flow
cytometer allows us to eliminate unwanted cells (like
erythrocytes) from the saved analysis
46. Thresholding (or contouring) on the
laser scanning cytometer
• On the laser scanning cytometer, any event that is above the threshold
(usually DNA fluorescence or sometimes forward scatter) is also
considered a “cell” and is also displayed for all parameters.
• The instrument marks such
events with a red “contour”,
which contains the amount
of fluorescent signal above
the threshold.
• Thresholding on the LSC is
therefore referred to as
“contouring”
47. Event-based contouring on the iCys
The red region is the event or thresholding contour.
It is analogous to “thresholding” or “discrimination”
on the flow cytometer. It defines the minimum
signal intensity that defines a cell. Everything else
below it is ignored. Like in cytometry, you need a
universal parameter (like scatter or DNA
luorescence) as the trigger for threshold contouring
The green region is the inte grating contour. You
can set this any number of pixels out from the
thresholding contour and measure the brightest
pixel (Max Pixel), or the total fluorescence
(Integral) within the green region.
The blue regions are the background contours.
You can also set these any number of pixels out
from the integrating contour, away from the cell.
The signal between them is interpreted as the
autofluorescence background, which is subtracted
from the other signals.
48.
49. Parameters studied by LSC
1. Integrated fluorescence intensity
2. The maximal intensity
3. The Integration area
4. The perimeter of the integration contour
(in micrometers).
5. The fluorescence intensity integrated
over the area of a torus of desired width
defined by the peripheral contour
located around (outside) of the primary
integration contour.
50. 6. The X-Y coordinates of maximal pixel
locating the measured object on of the
microscope stage.
7. The computer clock time at the moment
of measurement.
52. In Cytology
• It negates disadvantages of
Flow Image
cytmetry analysis
Requires Experience of
sufficient cytologist
amount needed
Verification
/reproducibility
not possible
Not for
adherent cells
54. Apoptosis studies
• Characterized by certain morphological
features e.g. nuclear fragmentation,
nuclear condensation
• Methods e.g. annexin V, loss of
transmembrane potential in
mitochondria, analysis of nuclear
fragmentation, alteration of DNA
condensation.
55.
56. Immunophenotyping of leucocyte
• Especially useful when amount of
obtained sample is minimum e.g.
neonate, critically ill patients
• Upto 5 flurochromes in < 15 microlit of
peripheral blood.
• Analysis by 2 methods
– Specific Immunostaining e.g. CD45
– DNA staining with 7-AAD- difference in
intensity with different leucocytes
57.
58. Ploidy and DNA index
• Anueploid number characteristic of a
malignant cell e.g. Gastric, colon,
kidney, head and neck etc.
• Prognostic marker in several tumors
• LSC measures amount of DNA in cell
59. Tumor cells are
identified and
gated based
upon cytokeratin
staining.
To verify the morphology of the two
populations, cells can be restained
with Wright-Giemsa or H & E. Cells
from each region can be relocated
and visualized by the CompuSort™
process.
60. Bacterial detection
• detect live and dead
bacteria,
• estimate cell numbers,
and
• calculate live/dead
ratios.
• The methods are propidium iodide (PI), unable to
permeate the intact cell membrane
easier and more of a living cell, but does label dead
accurate than cells with red fluorescence.
SYTO® 16(Molecular Probes) will
traditional, manual label the nucleic acids of living cells
counts. with green fluorescence. Shown
here are E. coli bacteria captured
on a membrane and stained with PI
and SYTO 16
he TransCyt® filter has been plunged into the sample, it rotates at a high speed and facilitates cell and mucus dispersion. A vacuum is then applied to the filter, which collects cells on a 5 μm porosity membrane. A software program allows a homogeneous deposition of cells until saturation. The TransCyt filter is then inverted and a positive pressure allows cells to adhere to an electronegative slide. After insertion of another TransCyt filter and of another slide, the whole procedure may be repeated until the entire sample has been treated.
Image or scanning cytometry (IC) combines imaging and cytometric analysis in a single technology platform.Rather than randomly imaging an entire field (like a microscope does), it selects, images and measures cells that meet certain user-adjustable criterion (such as size or fluorescence). It is therefore not random, but event-based.It is closely related to conventional flow cytometry, which also analyzes individual cells that meet certain characteristics (and is also event-based). Both are therefore cytometric techniques Unlike conventional flow cytometry, IC usually analyzes cells fixed to a horizontal surface. However, the technology (light sources, detectors, etc.) is very analogous to traditional flow cytometry. With scanning cytometry, imagery becomes a parameter, and relates to the other parameters (scatter and fluorescence).
The microscope (Olympus Optical Co.) is the key part of the instrument, and it providesessential structural and optical components (Fig. 1). The specimen deposited on a micro-scope slide on the stage of the microscope is illuminated by laser beams that rapidly scan the slide. The beams from two lasers (argon ion and helium-neon) spatially merged by dichroic mirrors are directed onto the computer controlled oscillating (350 Hz) mirror, which reflects them through the epi-illumination port of the microscope and images through the objective lens onto the slide. The mirror oscillations cause the laser beams to sweep the area of micro-scope slide under the lens. The beam spot size varies depending on the lens magnification, from 2.5 (at 40x) to 10.0 m (at 10x). The slide, with its xy position monitored by sensors, is placed on the computer-controlled motorized microscope stage, which moves at 0.5 m steps per each laser scan, perpendicularly to the scan. Laser light scattered by the cells is imaged by the condenser lens and its intensity recorded by sensors. The specimen-emitted fluorescence is collected by the objective lens and part of it is directed to a charge-coupling device(CCD) camera for imaging. Another part is directed to the scanning mirror. Upon reflection,it passes through a series of dichroic mirrors and optical emission filters to reach one of the four photomultipliers. Each photomultipler records fluorescence at a specific wavelength range, defined by the combination of filters and dichroic mirrors. A light source, additional to the lasers, provides transmitted illumination to visualize the objects through an eyepiece or the CCD camera.
1. Integrated fluorescence intensity representing the sum of intensities of all pixels (pic-ture elements) within the integration contour area. The latter may be adjusted to a de-sired width with respect to the threshold contour (Fig. 2).2. The maximal intensity of an individual pixel within this area (maximal pixel).3. The integration area, representing the number of pixels within the integration contour.4. The perimeter of the integration contour (in micrometers).5. The fluorescence intensity integrated over the area of a torus of desired width definedby the peripheral contour located around (outside) of the primary integration contour.For example, if the integration contour is set for the nucleus, based on red fluorescence(DNA stained by propidium iodide, PI), then the integrated (or maximal pixel) greenfluorescence of FITC (fluoresceinisothiocyanate)-stained cytoplasm can be measuredseparately, within the integration contour (i.e., over the nucleus) and within the periph-eral contour (i.e., over the rim of cytoplasm of desired width outside the nucleus). Allabove values of fluorescence (1,2,4) are automatically corrected for background,which is measured outside the cell, within the background contour (Fig. 2).6. The Xy coordinates of maximal pixel locating the measured object on of the microscopestage. 7. The computer clock time at the moment of measurement.The measurements by LSC are relatively rapid; having optimal cell density on the slide upto 5000 cells can be measured per minute. The accuracy and sensitivity of cell fluorescence measurements by LSC are comparable to the advanced flow cytometers
Intensity of maximal pixel of fluorescence is a sensitive marker of local hypo- or hyper-chromicity within the cell, reflecting low or high concentration (density) of the fluorescent probe. One of the early applications of LSC along this line was to detect chromatin conden-sation. Namely, DNA in condensed chromatin, such as mitotic or apoptotic cells, shows in-creased stainability (per unit area of chromatin image) with most fluorochromes. Thus, evenwhen integrated fluorescence of the analyzed cells (representing their DNA content) is the same, the intensity of maximal pixel of the cell with condensed chromatin is higher than that of the cell with more diffuse chromatin structure. Maximal pixel of the DNA-associated fluorescence was used as a marker to distinguish mitotic and immediately post-mitotic G 1 cells from interphase cells [5,6]. Although mitotic cells can be recognized by FC using a variety of markers [reviewed in 7], the advantage of this approach by LSC is that a single fluorochromeis used to discriminate between G 1 vs S vs G 2 vs M phase cells.