Methods in Histology

InCheat Sheets / Histology

Briefly describe the stages of tissue preparation for routine histological staining
1. Fixation: The organ or tissue is fixed with 10% Neutral Buffered Formalin. This terminates metabolism, prevents autolysis, kills pathogens, and hardens the tissue for sectioning.
2. Dehydration: The tissue is dehydrated in ascending concentrations of alcohol up to 100% to remove water
3. Clearing: Alcohol is removed from the tissue using organic solvents such as xylol or toluol
4. Embedding: The tissue is embedded in liquid paraffin. This converts the tissue into a solid form that can be sectioned.
5. Sectioning: The tissue is cut using a microtome to provide very thin sections that can be viewed using a microscope
6. Staining: The tissue is stained using Hematoxylin and Eosin

Dr. Jeffrey Kalei

Author and illustrator for Hyperexcision. Interested in emergency room medicine. I have a passion for medical education and drawing.

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Biochemistry

Adrenocorticotrophic hormone	P: <80 ng/L
Alanine aminotransferase (ALT)	P: 5–35 U/L
Albumin	P: 35–50 g/L
Aldosterone	P: 100–500 pmol/L
Alkaline phosphatase	P: 30–130 U/L (adults)
α-Amylase	P: 0–180 IU/dL
α-Fetoprotein	S: <10 kU/L
Angiotensin II	P: 5–35 pmol/L
Antidiuretic hormone (ADH)	P: 0.9–4.6 pmol/L
Aspartate transaminase	P: 5–35 U/L
Bicarbonate	P: 24–30 mmol/L
Bilirubin	P: 3–17 μmol/L
BNP	P: <50 ng/L
C-reactive protein	P: <10 mg/L
Calcitonin	P: <0.1 mcg/L
Calcium (ionized)	P: 1.0–1.25 mmol/L
Calcium (total)	P: 2.12–2.60 mmol/L
Chloride	P: 95–105 mmol/L
Cholesterol	P: <5.0 mmol/L
VLDL	P: 0.128–0.645 mmol/L
LDL	P: <2.0 mmol/L
HDL	P: 0.9–1.93 mmol/L
Cortisol	P: AM 450–700 nmol/L, Midnight 80–280 nmol/L
Creatine kinase (CK)	P: ♂ 25–195 U/L, ♀ 25–170 U/L
Creatinine	P: 70–100 μmol/L
Ferritin	P: 12–200 mcg/L
Folate	S: 2.1 mcg/L
FSH	P/S: 2–8 U/L ♂ (luteal); >25 U/L menopause
Gamma-glutamyl transpeptidase	P: ♂ 11–51 U/L, ♀ 7–33 U/L
Glucose (fasting)	P: 3.5–5.5 mmol/L
Growth hormone	P: <20 mu/L
HbA1C (DCCT)	B: 4–6%
HbA1C (IFCC)	B: 20–42 mmol/mol
Iron	S: ♂ 14–31 μmol/L, ♀ 11–30 μmol/L
Lactate	P: Venous 0.6–2.4 mmol/L, Arterial 0.6–1.8 mmol/L
Lactate dehydrogenase (LDH)	P: 70–250 U/L
Lead	B: <1.8 mmol/L
LH	P: 3–16 U/L (luteal)
Magnesium	P: 0.75–1.05 mmol/L
Osmolality	P: 278–305 mosmol/kg
Parathyroid hormone (PTH)	P: 0.8–8.5 pmol/L
Potassium	P: 3.5–5.3 mmol/L
Prolactin	P: ♂ <450 U/L, ♀ <600 U/L
PSA	P: 0–4 mcg/mL
Protein (total)	P: 60–80 g/L
Red cell folate	B: 0.36–1.44 μmol/L
Renin	P: Erect 2.8–4.5, Recumbent 1.1–2.7 pmol/mL/h
Sodium	P: 135–145 mmol/L
TBG	P: 7–17 mg/L
TSH	P: 0.5–4.2 mU/L
Thyroxine (T4)	P: 70–140 nmol/L
Free T4	P: 9–22 pmol/L
Total iron-binding capacity	S: 54–75 μmol/L
Triglycerides (fasting)	P: 0.50–2.3 mmol/L
Triiodothyronine (T3)	P: 1.2–3.0 nmol/L
Troponin T	P: <0.1 mcg/L
Urate	P: ♂ 210–480 μmol/L, ♀ 150–390 μmol/L
Urea	P: 2.5–6.7 mmol/L
Vitamin B12	S: 0.13–0.68 nmol/L
Vitamin D	S: 50 nmol/L

Arterial Blood Gases

pH	7.35–7.45
PaCO₂	4.7–6.0 kPa
PaO₂	>10.6 kPa
Base excess	±2 mmol/L

Urine

Cortisol (free)	<280 nmol/24h
Hydroxyindole acetic acid	16–73 μmol/24h
Hydroxymethylmandelic acid	16–48 μmol/24h
Metanephrines	0.03–0.69 μmol/mmol creatinine
Osmolality	350–1000 mosmol/kg
17-Oxogenic steroids	♂ 28–30 μmol/24h, ♀ 21–66 μmol/24h
17-Oxosteroids (neutral)	♂ 17–76 μmol/24h, ♀ 14–59 μmol/24h
Phosphate (inorganic)	15–50 mmol/24h
Potassium	14–120 mmol/24h
Protein	<150 mg/24h
Protein/creatinine ratio	<3 mg/mmol
Sodium	100–250 mmol/24h

Haematology

WCC	4.0–11.0 ×10⁹/L
Red cell count	♂ 4.5–6.5 ×10¹²/L, ♀ 3.9–5.6 ×10¹²/L
Haemoglobin	♂ 130–180 g/L, ♀ 115–160 g/L
PCV / Haematocrit	♂ 0.4–0.54 L/L, ♀ 0.37–0.47 L/L
MCV	76–96 fL
MCH	27–32 pg
MCHC	300–360 g/L
RDW	11.6–14.6%
Neutrophils	2.0–7.5 ×10⁹/L (40–75%)
Lymphocytes	1.0–4.5 ×10⁹/L (20–45%)
Eosinophils	0.04–0.44 ×10⁹/L (1–6%)
Basophils	0–0.10 ×10⁹/L (0–1%)
Monocytes	0.2–0.8 ×10⁹/L (2–10%)
Platelet count	150–400 ×10⁹/L
Reticulocyte count	0.8–2.0% / 25–100 ×10⁹/L
ESR	Depends on age
Prothrombin time	10–14 s
APTT	35–45 s

Paediatric

Pulse Rate (beats per minute)
Neonate (<1 month)	140–160
Infant (<1 year)	120–140
1–5 years	110–130
5–12 years	80–120
>12 years	70–100
Temperature
Normal range	36.6–37.2 °C
38.0 °C	Fever (infant)
38.5 °C	Fever (>3 months)
Respiratory Rate (tachypnoea)
Neonate (0–2 months)	≥60 breaths/min
2–12 months	≥50 breaths/min
1–5 years	≥40 breaths/min
>5 years	≥30 breaths/min
Blood Pressure (mmHg)
Term	65/45
1 year	75/50
4 years	85/60
8 years	95/65
10 years	100/70
Head Circumference
Term	35 cm ± 2
1 year	+1 cm/month
2–3 years	+2 cm/year
4–5 years	+1 cm/year
5 years	~53 cm
10 years	56–57 cm
Height
Term	52–55 cm
1 year	75 cm
2–12 years	6a + 77 cm
Weight Formulas
3–12 months	(a + 9)/2 kg
1–6 years	2a + 8 kg
>6 years	(7a − 5)/2 kg
Developmental Milestones
Social smile	1.5 months
Head control	4 months
Sits unsupported	7 months
Crawls	10 months
Stands unsupported	10–12 months
Walks	12–13 months
Talks	18 months
MUAC (6 months–5 years)
Obese	>17.5 cm
Normal	13.5–17.4 cm
At risk	12.5–13.4 cm
Moderate malnutrition	11.5–12.4 cm
Severe malnutrition	<11.5 cm
Blood Transfusion
Whole blood	20 mL/kg
pRBC	10 mL/kg
Formula	6 × (weight in kg) × Hb deficit
Normal Paediatric Lab Values
Haemoglobin (g/dL)	Hematocrit (%)
Term newborn: 13–20	40–58
1 month: 11–18	32–54
2 months: 10–15	28–44
1–2 years: 10–13	32–40
>2 years: 11–14	33–42
CSF WBC (/mm³)
Term newborn	0–25
>2 weeks	0–5

Dr. Jeffrey Kalei

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