17_HST110 Introduction to the Histology of the Kidney-The Basic Techniques Flashcards
What stains are used in light microscopy analysis of morphological feature of the kidney?
H&E, PAS, silver methenamine (Jones), trichrome, (Congo red; von Kossa, alizarin red)
Under immunohistochemistry, what proteins are tagged in immunofluorescence?
IgG, IgA, IgM, kappa, lambda, C3, C1q, C4d, albumin, fibrin, (protein A, transthyretin, collagen)
Under immunohistochemistry morphological techniques, what does IMPOX look for?
viruses (BK, CMV, adenovirus), lymphocyte and other cell markers, lysozyme, hemoglobin, myoglobin, etc.
What does one look for using electron microscopy of renal tissues?
podocyte integrity, lysosomal inclusions, location of EDD, substructure in EDD, fibrillary material and measurements, texture and thickness of the BM, viral particles, mitochondrial changes, etc.
What fixative is used for light microscopy preservation and preparation before paraffin embedding?
Formaldehyde solution
What solution is used to prevent protease-dependent protein disturbance for immunofluorescence microscopy?
Michel’s solution
What fixative solution is used for electron microscopy preservation and preparation?
Paraformaldehyde/glutaraldehyde solution (Karnovsky’s Fixative)
PAS (periodic acid Schiff) is a histochemical technique detecting sugar moieties to highlight (X). PAS stains glycosylated proteins (Y)
X = basement membrane Y = purple
What cellular features need to be considered in glomerular pathology by LM?
Type of cells (mononuclear, PMN, Eos etc.)
Location of cells (tuft vs. Bowman’s space
Capillaries vs. mesangial space)
Intensity and frequency
What spaces need to be considered in glomerular pathology by LM?
Capillary lumen (open, occluded, collapsed) Bowman’s space (open, distended, cells)
What matrix features need to be considered in glomerular pathology by LM?
Basement membrane (thickness, “spikes” and “craters”, duplicated (double contours)
Mesangial matrix (nodules, PAS-positivity, Congo red); scar collagen
What is the definition of a focal issue?
A subset of elements is involved (<50%)
What is the definition of a diffuse issue?
All elements are involved (>50%)
What is the definition of a global issue?
The entire element (e.g glomerulus) is involved
What is the definition of a segmental issue?
A segment or sector of the element (e.g. glomerulus) is involved
What does endocapillary mean?
Affecting the tuft
What does extracapillary mean?
Affecting the Bowman’s space
What does proliferative mean?
Increase in number of cells, exudative (inflammatory)
What does membranous mean in the context of renal pathophysiology?
Thickened capillary walls (“spikes”)
What does membranoproliferative mean in the context of renal pathophysiology?
Thickened capillary wall (double contours) and increase in the number of cells
The hematoxylin and eosin stains nuclei in (X) and the cytoplasm and various proteins in (Y)
X = blue Y = red-pink-orange
Hemoglobin is not strongly stained by (X) (RBCs not strongly glycosylated)
X = PAS
Lysosomal accumulation of proteins (as in (X) disease) can lead to massive vacuolization of the (Y) since they are never regenerated (other cells affected, but can be renewed before extensive vacuoles accumulate)
X = Fabry Y = podocytes
Glomerulitis is (X) of the glomerulus, either segmental or global
X = inflammation
A global glomerulitis is mostly due to infilration of (X) inflammatory cells exhibiting a lobular pattern of injury
X = mononuclear
A cellular crescent, focal fibrinoid necrosis, and shrunken glomerulus can result in the case of inflammation due to the accumulation of (X) and parietal epithelial cells in Bowman’s space
X = macrophages/monocytes
Nodular accumulation of PAS-positive matrix in the mesangium is a classic presentation in (X)
X = DM (Diabetes Mellitus)
Amyloid deposits stain weekly with PAS and exhibit a characteristic (X) when stained with Congo red and viewed under polarized light
X = apple green birefringence
(X) are commonly seen in Membranoproliferative glomerulonephritis, where a second basement membrane is deposited after injury
X = Double contours
What type of collagen is considered the scar collagen seen in the healing of a focal necrotizing lesion?
Collagen I (Lightly staining in PAS)
For IF microscopy section preparation, the tissue is not fixed. Instead, it is (X)
X = snap frozen
What are the major elements to consider in glomerular pathology by IFM?
The antibodies used are chain-specific The distribution of the deposits Type of deposits Location of the deposits Intensity of staining (0 to 3+ or 4+)
What chain -specific antibodies are considered in IFM?
anti-IgG=anti-g heavy chain; etc.
What are potential locations of deposits seen in IFM?
peripheral capillary wall, mesangium, capillary lumen, Bowman space, tubular basement membranes, vascular wall, nuclear staining
What cells produce the kappa or lambda light chains which have affinity for the glomerulus?
B cells and plasma cells
Tissue is embedded in hard (X) to achive ultra-thin sections of the tissue on the ultramicrotome for EM
X = epoxy resin
High protein content can be seen as (X) regions under EM. E.g. RBCs
electron dense
(X) on the endothelium in the kidney is much higher than in other organs with lower perfusion/gram (such as the brain). Higher pressure also seen in this system. Thus, the kidney expresses thrombotic and cardiovascular diseases long before other organs become clinically involved
X = Shear stress
What type of hemoglobin forms the fibrils seen in sickle cell anemia?
Hemoglobin S
Lyme disease exhibits (X) in the glomerular basement membrane
X = double contours
Immune complex accumulation on the luminal side of glomerular capillaries is a classic presentation of (X)
X = serum sickness
Deposits in monoclonal IgG/kappa deposition disease appear almost (X)
X = crystalline
AL (Lambda-derived) amyloidosis shows podocytes being pushed aside by (X) of fibrils, in appearance similar to crystal growth
X = spicules
(X) can be used to analyze local protein content to discover and/or back-calculate abnormal quantities, types, or localizations
X = Laser microdissection + peptide extraction + tandem spectrometry
