Laboratory0112

28/07/2025

Fertility Test (Microscopic Examination of Semen)

1. Objective:

The objective of the fertility test was to assess male fertility by examining semen for s***m count, motility, morphology, and other parameters under a microscope.

2. Principle:

The principle of the test was based on microscopic analysis of semen to evaluate the quality and quantity of s***matozoa. This helped determine the potential fertility of a male.

3. Materials:

Clean, sterile semen collection container

Glass slides and coverslips

Micropipette or dropper

Light microscope (with 40x magnification)

Timer or stopwatch

Staining reagents (e.g., Eosin-Nigrosin for morphology)

Incubator (37°C)

Personal protective equipment (PPE)

4. Procedure (Microscopic):

1. The semen sample was collected after 2–7 days of abstinence in a sterile container.

2. It was allowed to liquefy at room temperature for 30 minutes.

3. A drop of semen was placed on a glass slide and covered with a coverslip.

4. Under the microscope, s***m motility was first observed at 40x magnification.

5. Another slide was prepared and stained for assessing morphology and counted under oil immersion (100x).

6. S***m concentration was calculated using a Neubauer counting chamber if needed.

7. Volume, viscosity, and appearance were also noted.

5. Result:

A normal semen sample showed >15 million s***m/mL, with at least 40% motile and >4% normal morphology.

Abnormal results included low s***m count (oligos***mia), poor motility (asthenozoos***mia), or no s***m (azoos***mia).

6. Uses:

It was used to assess male fertility in couples facing infertility issues.

It helped diagnose reproductive tract infections, hormonal imbalances, or genetic disorders affecting s***m production.

7. Consultation:

Based on the results, patients were advised to consult a fertility specialist or urologist. Further hormonal tests, imaging, or genetic counseling were recommended if abnormalities were detected.................
Female Fertility Test

1. Objective:

The objective of the female fertility test was to assess the reproductive health of a woman by evaluating ovulation, cervical mucus, and hormonal levels.

2. Principle:

The principle was based on detecting signs of ovulation and the body's readiness for conception. Microscopic analysis of cervical mucus and hormone testing (like LH, FSH, and estrogen) provided insights into the woman’s fertility status.

3. Materials:

Sterile swab or speculum

Glass slides and coverslips

Microscope

Staining solution (optional)

Hormonal assay kits (for LH, FSH, prolactin, estrogen)

Basal body temperature (BBT) chart

Ovulation prediction kit (optional)

4. Procedure (Microscopic):

1. A cervical mucus sample was collected around the mid-cycle (Day 12–16).

2. A drop of mucus was placed on a slide and covered with a coverslip.

3. The sample was observed under a microscope for fern-like crystallization, indicating the presence of estrogen and ovulation.

4. Hormonal tests were performed using blood samples to check levels of LH, FSH, estradiol, and progesterone.

5. Basal body temperature charts were reviewed to confirm ovulatory patterns.

5. Result:

Normal results showed fern pattern in cervical mucus and hormone levels within normal ovulatory ranges.

Abnormal findings included absence of ferning, low progesterone (indicating anovulation), or high FSH (suggesting poor ovarian reserve).

6. Uses:

It was used to identify causes of female infertility.

Helped determine ovulation status and hormonal imbalances.

Guided treatment in cases like polycystic ovarian syndrome (PCOS), thyroid issues, or early menopause.

7. Consultation:

Based on the results, women were referred to a gynecologist or reproductive endocrinologist. Additional imaging tests like transvaginal ultrasound or hysterosalpingography (HSG) were suggested if needed.

20/06/2025

Job opportunities

09/06/2025

Crossmatch Test (Major & Minor) in a standard lab report format:

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1. Objective

The objective of the test was to detect any incompatibility between the donor's and recipient's blood, ensuring safe blood transfusion by performing both major and minor crossmatch tests.

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2. Principle

The principle of the crossmatch test was based on detecting agglutination or hemolysis that occurred when donor red blood cells and recipient serum (major) or recipient red blood cells and donor serum (minor) were mixed. Any incompatibility indicated the presence of antibodies that could cause transfusion reactions.

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3. Materials

Donor blood sample

Recipient blood sample

Centrifuge

Test tubes

Normal saline

Anti-human globulin (AHG) reagent

Incubator

Pipettes

Slide or glass plates

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4. Procedure

Major Crossmatch:

1. Donor red cells were washed with normal saline and a 2–5% cell suspension was prepared.

2. Two drops of recipient serum and one drop of donor cell suspension were mixed in a clean test tube.

3. The mixture was incubated at 37°C for 15–30 minutes.

4. After incubation, the tube was centrifuged, and agglutination was observed.

5. The indirect Coombs test (AHG phase) was also performed to detect incomplete antibodies.

Minor Crossmatch:

1. Recipient red cells were washed and a 2–5% cell suspension was prepared.

2. Two drops of donor serum and one drop of recipient cell suspension were mixed in another test tube.

3. Similar steps of incubation, centrifugation, and observation were followed as in the major crossmatch.

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5. Result

Agglutination or hemolysis indicated an incompatible crossmatch.

If no agglutination or hemolysis was observed in both major and minor crossmatch, the result was compatible.

If agglutination or hemolysis was present in either test, it was incompatible and the donor blood was deemed unsafe for transfusion.

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6. Uses

The test was used to prevent transfusion reactions.

It confirmed ABO and Rh compatibility.

It was used to detect unexpected antibodies in the recipient’s serum.

It ensured the safety of blood transfusions in clinical settings.

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7. Conclusion

The crossmatch test successfully determined the compatibility between donor and recipient blood. Performing both major and minor crossmatches helped identify any immune reaction risks and ensured safe transfusion practices.

08/06/2025

Antibody Screening & Identification Panels test:

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🧪 Antibody Screening & Identification Panels (in Immunohematology)

1. Objective:
To detect and identify unexpected antibodies in a patient’s serum that may react with red blood cell antigens, especially before transfusion.

2. Principle:
Patient serum is mixed with reagent red cells of known antigen profiles. Agglutination indicates the presence of antibodies. Identification involves comparing reaction patterns with known antigen matrices.

3. Materials:

Patient serum/plasma

Screening and identification red cell panels

LISS or saline

Antiglobulin reagent (Coombs reagent)

Test tubes or gel cards

Centrifuge, incubator

4. Procedure:

1. Mix patient serum with screening cells → incubate.

2. Wash to remove unbound antibodies.

3. Add antihuman globulin → check for agglutination.

4. If positive, proceed to identification with an extended red cell panel.

5. Compare agglutination pattern to known antigen profiles.

5. Result Interpretation:

No agglutination: No detectable antibodies.

Agglutination with specific cells: Suggests presence of particular antibody (e.g., anti-D, anti-K, etc.).

6. Clinical Significance:

Crucial in transfusion safety.

Prevents hemolytic transfusion reactions.

Used in prenatal testing for HDFN (Hemolytic Disease of the Fetus and Newborn).

08/06/2025

🧪 Rh Antibody Titration Test

1. Objective:
To determine the concentration (titer) of anti-D (Rh) antibodies in a patient's serum—especially in Rh-negative pregnant women—to assess risk of Hemolytic Disease of the Fetus and Newborn (HDFN).

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2. Principle:
Serial dilutions of the patient’s serum are reacted with Rh-positive red cells. The highest dilution at which agglutination still occurs is the antibody titer. This provides a quantitative estimate of antibody strength.

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3. Materials Required:

Patient's serum

Rh(D) positive red blood cells (usually R1R1)

Phosphate-buffered saline (PBS)

2–5% red cell suspension

Incubator at 37°C

Centrifuge

Test tubes

Antihuman globulin (AHG or Coombs reagent)

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4. Procedure:

1. Prepare serial twofold dilutions of the patient’s serum (e.g., 1:1 to 1:1024).

2. Add equal volume of 2–5% Rh(D)-positive RBC suspension.

3. Incubate at 37°C for 30–60 minutes.

4. Wash cells 3–4 times with PBS.

5. Add AHG reagent, centrifuge, observe for agglutination.

6. The highest dilution showing 1+ agglutination or stronger is recorded as the titer.

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5. Result Interpretation:

Titer ≥16–32: Potential risk of HDFN, requires close monitoring.

Titer rising over time: Indicates active alloimmunization.

Stable or low titer: Lower risk, but monitor.

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6. Clinical Relevance:

Helps guide obstetric management in Rh-incompatible pregnancies.

Determines need for intrauterine transfusion or early delivery.

07/06/2025

Direct and Indirect Coombs Test (Antiglobulin Test) o
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1. Objective

The objective of this test was to detect antibodies that act against red blood cells (RBCs), either attached to the RBC surface (Direct Coombs) or circulating freely in serum (Indirect Coombs), to diagnose conditions such as autoimmune hemolytic anemia, hemolytic disease of the newborn, or for cross-matching in transfusions.

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2. Principle

The Coombs test was based on the principle of antigen-antibody reaction.

Direct Coombs Test (DAT): Detected antibodies or complement bound in vivo to RBCs.

Indirect Coombs Test (IAT): Detected free antibodies in vitro in the serum that could bind to antigens on donor RBCs.
Both tests used anti-human globulin (AHG) reagent to cause agglutination if sensitization was present.

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3. Materials Used

Patient’s blood sample (for DAT)

Patient’s serum and donor RBCs (for IAT)

Coombs reagent (AHG)

Test tubes

Centrifuge

Normal saline

Pipettes

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4. Procedure

Direct Coombs Test (DAT):

1. Patient’s RBCs were washed 3 times with saline to remove unbound proteins.

2. 1–2 drops of AHG reagent were added.

3. The tube was gently mixed and centrifuged.

4. The RBCs were examined for agglutination.

Indirect Coombs Test (IAT):

1. Patient’s serum was mixed with reagent RBCs (with known antigens).

2. The mixture was incubated at 37°C for 15–30 minutes.

3. The cells were washed with saline to remove unbound antibodies.

4. AHG was added, centrifuged, and checked for agglutination.

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5. Result Interpretation

Positive: Visible agglutination occurred, indicating the presence of antibodies bound to RBCs (DAT) or free antibodies capable of sensitizing RBCs (IAT).

Negative: No agglutination was observed.

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6. Use Cases

Direct Coombs: Autoimmune hemolytic anemia, hemolytic disease of the newborn.

Indirect Coombs: Pre-transfusion testing, Rh incompatibility screening in pregnancy.

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