4 - CLINICAL MICROSCOPY

UC-3500

Urine Analyzers (Main Machines)

Fully Automated Urine Chemistry Analyzer

Reflectance Photometry with Colour CMOS Sensor

Urine Analyzers (Main Machines)

UC-3500

Principle

Glucose

Ketone

Bilirubin

Specific gravity

Protein

pH

Urobilinogen

Nitrite

Leukocyte esterase

Blood

Urine Analyzers (Main Machines)

UC-3500

Parameters (10)

Low

High

Urine Analyzers (Main Machines)

UC-3500

• Quality Control

  • 2 levels: _ (white cap) and _ (yellow cap)

  • Thrice a day: 6AM, 2PM, and 10PM

UF-4000

Urine Analyzers (Main Machines)

Urine Sediment Testing

Flow Cytometry with Hydrodynamic Focusing

Urine Analyzers (Main Machines)

UF-4000

Principle

RBCs

WBCs

Epithelial Cells

Casts

Bacteria

Urine Analyzers (Main Machines)

UF-4000

5 Main Parameters:

Low

High

Urine Analyzers (Main Machines)

UF-4000

• Quality Control

  • 2 levels: _ (blue) and _ (red)

  • Thrice a day: 6AM, 2PM, 10PM

DCA Vantage

Urine Analyzers (Main Machines)

Test: Urine Microalbumin-Creatinine Ratio

Spectrophotometry

Urine Analyzers (Main Machines)

DCA Vantage

Principle: Analyzes the intensity of light that is transmitted to the cartridge optical window and reports the results in clinically meaningful units

Clinitek Advantus

Urine Analyzers (Main Machines)

• Test/s:

  • Urine Reagent Strip Testing

  • Urine Free Hemoglobin

  • Urine Myoglobin

  • Serum Ketone

Reflectance Photometry

Urine Analyzers (Main Machines)

Clinitek Advantus

Principle:

Fecal Immunochemical Test (FIT)

Most Commonly Performed Special Tests

Often used to detect bleeding in the digestive tract which has no other signs or symptoms

More specific to finding blood coming from the lower gastrointestinal tract than the FOBT

8 drops (alternating manner) for 5 minutes

3 drops for 5 minutes

Most Commonly Performed Special Tests

Fecal Immunochemical Test (FIT)

1. Insure One: ?

2. Hemoccult ICT: ?

4 drops for 3 minutes

3 drops for 3 minutes

Most Commonly Performed Special Tests

Pregnancy Test Kits

1. Wondfo: ?

2. Rightsign: ?

3 drops for 10 minutes

Most Commonly Performed Special Tests

CERTEST Kits

? for ALL CERTEST test kits

Coloured chromatographic immunoassay

Most Commonly Performed Special Tests

CERTEST Kits

Principle?

H. pylori stool antigen

Most Commonly Performed Special Tests

CERTEST Kits

• Coloured chromatographic immunoassay for the qualitative detection of Helicobacter pylori in stool samples

• The importance of H. pylori testing has increased greatly since the strong correlation between the presence of bacteria and confirmed gastrointestinal diseases (stomach and duodenum) like gastritis, peptic ulcer disease and gastric carcinoma

Fecal Calprotectin Semi-Quantitative

Most Commonly Performed Special Tests

CERTEST Kits

• Coloured chromatographic immunoassay for the semi-quantitative detection of human calprotectin (hCp) in stool samples that may reflect gastrointestinal inflammation caused by several pathologies (inflammatory bowel disease, colorectal cancer, and some enteropathies)

Fecal Occult Blood (FOBT) Semi-Quantitative

Most Commonly Performed Special Tests

CERTEST Kits

• Coloured chromatographic immunoassay for the semi-quantitative determination of human haemoglobin (hHb) in stool samples

Hemascreen Fecal Occult Blood (FOBT) Qualitative

Most Commonly Performed Special Tests

• Stored at room temperature

• Rapid, qualitative method for detecting occult blood in the stool which may be indicative of asymptomatic gastrointestinal diseases such as colorectal cancer, polyps, or colitis

2 drops of buffer for each window for 30 seconds

Most Commonly Performed Special Tests

Hemascreen Fecal Occult Blood (FOBT) Qualitative

• Drops & minutes

Parascreen: Malaria Antigen Test Kit

Most Commonly Performed Special Tests

• Rapid, qualitative, two site sandwich immunoassay utilizing whole blood for the detection of P. falciparum specific histidine rich protein-2 (Pf. HRP-2) and Pan malaria specific pLDH

• The test may also be used for the differentiation of P. falciparum and other material species (P. malariae, P. vivax, P. ovale, and P. knowlesi) and for the follow up of antimalarial therapy in whole blood samples

sandwich immunoassay

5 uL of blood with 2 drops of buffer for 20 minutes

Most Commonly Performed Special Tests

Parascreen: Malaria Antigen Test Kit

• Principle

• Drops and minutes

Operon: Simple H. pylori

Most Commonly Performed Special Tests

• Simple/Stick H.pyl* and Simple H.pyl-Tf are non-invasive rapid tests that can identify both active H. pylori infection and the presence of bleeding in the gastrointestinal tract quickly, safely, easily, affordably and reliably

4 drops for 15 minutes

Most Commonly Performed Special Tests

Operon: Simple H. pylori

• Drops & minutes

Operon: Simple Calprotectina

Most Commonly Performed Special Tests

• immunochromatographic assay is a rapid test for the semi-quantitative detection of human Calprotectin in fecal samples as an indicator of the gastrointestinal inflammation present in various pathologies (ulcerative colitis, Crohn’s disease, colorectal cancer)

immunochromatographic assay

Most Commonly Performed Special Tests

Operon: Simple Calprotectina

• Principle

MRP8 and MRP14

neutrophil

3 drops for 10 minutes

Most Commonly Performed Special Tests

Operon: Simple Calprotectina

• Calprotectin is a heterocomplex with antimicrobial properties comprised of two calcium-binding proteins (?) present in the cytoplasm of neutrophils

• The presence of calprotectin in feces is a consequence of _ migration into the gastrointestinal tissue due to an inflammatory process

• Drops and minutes

Wondfo: Fecal Occult Blood (FOBT) Semi-Quantitative

Most Commonly Performed Special Tests

• _ test uses immunochromatographic sandwich method, which employs two specific monoclonal antibodies to selectively identify hemoglobin in test samples

• It is a rapid immunochromatographic direct binding test for the visual detection of hemoglobin in fecal samples

immunochromatographic sandwich method

3 drops for 10 minutes

Most Commonly Performed Special Tests

Wondfo: Fecal Occult Blood (FOBT) Semi-Quantitative

• Principle

• Drops & minutes

Brown, Yellow Brown, Light Brown, Dark Brown, Greenish Brown, Greenish Black, Reddish

Hard, Formed, Soft, Mushy, Loose, Watery

Fecalysis

• Examine the gross appearance of the sample according to:

  • Color: (8)

  • Consistency: (6)

  • Mucus: Present (+) or Absent (-)

NSS

Lugol’s iodine

LPO, HPO

Fecalysis

• Procedure:

1. Get 1 slide. Using a marker, Write the patient’s initial at the side of the glass slide

2. Place 1 drop of _ on one part of the slide and another drop of _ _ on the area opposite to NSS

3. With an applicator stick, poke at various parts of the stool sample. Make sure to touch the portions with blood or mucus

4. Make a smooth uniform emulsion in both solutions

5. Place the coverslips in both emulsion (avoid air bubbles)

6. Examine both emulsions under _ to screen and detect the presence of helminth eggs and larvae. View under _ for greater morphologic details and better identification

Entamoeba coli

Entamoeba histolytica

Blastocystis hominis

Giardia lamblia cyst and trophozoite

Calcium Oxalate

Amorphous Urates & Amorphous Phosphates

Triple Phosphate

Ammonium Biurate

Hyaline Cast

Fine Granular Cast

Coarse Granular Cast

Mucus Threads

Sperm

RBCs

WBCs

Fat Globules

Yeast Cells

Budding Yeast Cells

Yeast Cells with Hyphae

8 drops, 5 mins (alt manner)

3 drops, 5 minutes

Special Tests
Test	Principle	Drops and Reading Time	Detects:
Fecal Immunochemical Tests (FIT)
Insure One		?	Detects bleeding in the gastrointestinal tract
Hemoccult ICT		?

4 drops, 3 minutes

3 drops, 3 minutes

Pregnancy Test Kits
Wondfo		?
Rightsign		?

Coloured Chromatographic Immunoassay

3 drops, 10 minutes

CERTEST KITS
H. pylori stool antigen	?	?	H. pylori – gastritis, peptic ulcer, gastric carcinoma
Fecal Calprotectin Semi-Quantitative			Human calprotectin – inflammatory bowel disease, colorectal cancer, enteropathies
Fecal Occult Blood (FOBT) Semi-Quantitative			Human hemoglobin

2 drops for each window, 30 secs

Sandwich Immunoassay

5 uL of blood w/ 2 drops of buffer for 10 minutes

4 drops, 15 minutes

Other Test Kits
Hemascreen Fecal Occult Blood (FOBT) Qualitative		?	Colorectal cancer, polyps, or colitis
Parascreen: Malaria Antigen Test Kit	?	?	P. falciparum (histidine rich protein-2) Pan malaria (pLDH)
Operon: Simple H. pylori		?	H. pylori infection
Operon: Simple Calprotectin	Immunochromatographic Assay	3 drops, 10 minutes	HCP – ulcerative colitis, Crohn’s Disease, colorectal cancer MRP8, MRP14
Wondfo: Fecal Occult Blood (FOBT) Semi-Quantitative	Immunochromatographic Sandwich Method	3 drops, 10 minutes	Visual detection of hemoglobin in fecal samples

Immunochromatographic Assay

3 drops, 10 minutes

Immunochromatographic Sandwich Method

3 drops, 10 minutes

Other Test Kits
Hemascreen Fecal Occult Blood (FOBT) Qualitative		2 drops for each window, 30 secs	Colorectal cancer, polyps, or colitis
Parascreen: Malaria Antigen Test Kit	Sandwich Immunoassay	5 uL of blood w/ 2 drops of buffer for 10 minutes	P. falciparum (histidine rich protein-2) Pan malaria (pLDH)
Operon: Simple H. pylori		4 drops, 15 minutes	H. pylori infection
Operon: Simple Calprotectin	?	?	HCP – ulcerative colitis, Crohn’s Disease, colorectal cancer MRP8, MRP14
Wondfo: Fecal Occult Blood (FOBT) Semi-Quantitative	?	?	Visual detection of hemoglobin in fecal samples

Total Cell Count

Differential Count

Additional Notes

• Body Fluids Cell Count & Gross Examination

  • _ _ _ – uncentrifuged

  • _ _ – centrifuged

NSS

Glacial Acetic Acid

Crystal Violet

Additional Notes

• Body Fluids Cell Count & Gross Examination

  • Reagents used:

    • _

    • _ _ _ – lyses RBC

    • _ _

NSS

glacial acetic acid

crystal violet

Additional Notes

Body Fluids Cell Count & Gross Examination

• Total Cell Count – _

• All body fluids – _ _ _

• Synovial fluid – _ _ (prevents mucin clot and clumping)

double sequential enzymatic reaction

Additional Notes

Reagent Strip Parameters:

Glucose

Diazo reaction

Additional Notes

Reagent Strip Parameters:

Bilirubin

sodium nitroprusside reaction

Additional Notes

Reagent Strip Parameters:

Ketone

pKa change of polyelectrolyte

Additional Notes

Reagent Strip Parameters:

Specific gravity

protein error of indicators

Additional Notes

Reagent Strip Parameters:

Protein

double indicator system

Additional Notes

Reagent Strip Parameters:

pH

Greiss reaction

Additional Notes

Reagent Strip Parameters:

Nitrite

pseudoperoxidase activity of Hgb

Additional Notes

Reagent Strip Parameters:

Blood

leukocyte esterase activity

Additional Notes

Reagent Strip Parameters:

Leukocyte esterase

Ehrlich reaction

Additional Notes

Reagent Strip Parameters:

Urobilinogen

Uric Acid

Acidic

_ _

_

Rhombic, four-sided flat plates (whetstones), wedges, rosette

Amorphous Urates

Acidic

?

?

Yellow-brown granules Refrigeration: Pink sediment

Amorphous Phosphate

Alkaline

Refrigeration: White sediment

Calcium Oxalate

Acidic/Alkaline

Dihydrate: Octahedral envelope Monohydrate: Dumbbell-shaped (ethylene glycol “antifreeze” poisoning)

Calcium Phosphate

Alkaline

Thin prism in rosette form

Triple Phosphate

Alkaline

AKA “Ammonium Magnesium Phosphate” or “Struvite” Coffin lid

Ammonium Biurate

Alkaline

Thorny apples Most often encountered in old specimen

Calcium Carbonate

Acidic/Alkaline

Dumbbell or spherical shape