Abstract
Immunohistochemical and exvivo anatomical studies have provided many glimpses of the variety, distribution, and signaling components of vertebrate retinal neurons. The beauty of numerous images published to date, and the qualitative and quantitative information they provide, indicate that these approaches are fundamentally useful. However, obtaining these images entailed tissue handling and exposure to chemical solutions that differ from normal extracellular fluid in composition, temperature, and osmolarity. Because the differences are large enough to alter intercellular and intracellular signaling in neurons, and because retinae are susceptible to crush, shear, and fray, it is natural to wonder if immunohistochemical and anatomical methods disturb or damage the cells they are designed to examine. Tissue fixation is typically incorporated to guard against this damage and is therefore critically important to the quality and significance of the harvested data. Here, we describe mechanisms of fixation; advantages and disadvantages of using formaldehyde and glutaraldehyde as fixatives during immunohistochemistry; and modifications of widely used protocols that have recently been found to improve cell shape preservation and immunostaining patterns, especially in proximal retinal neurons.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 181-202 |
| Number of pages | 22 |
| Journal | Progress in Retinal and Eye Research |
| Volume | 48 |
| DOIs | |
| State | Published - Sep 1 2015 |
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Keywords
- Antigen retrieval
- Fixatives
- Formaldehyde
- Glutaraldehyde
- PH
- Quenching
- Retina
- Sucrose
ASJC Scopus subject areas
- Sensory Systems
- Ophthalmology
Cite this
Fixation strategies for retinal immunohistochemistry. / Stradleigh, Tyler W.; Ishida, Andrew.
In: Progress in Retinal and Eye Research, Vol. 48, 01.09.2015, p. 181-202.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Fixation strategies for retinal immunohistochemistry
AU - Stradleigh, Tyler W.
AU - Ishida, Andrew
PY - 2015/9/1
Y1 - 2015/9/1
N2 - Immunohistochemical and exvivo anatomical studies have provided many glimpses of the variety, distribution, and signaling components of vertebrate retinal neurons. The beauty of numerous images published to date, and the qualitative and quantitative information they provide, indicate that these approaches are fundamentally useful. However, obtaining these images entailed tissue handling and exposure to chemical solutions that differ from normal extracellular fluid in composition, temperature, and osmolarity. Because the differences are large enough to alter intercellular and intracellular signaling in neurons, and because retinae are susceptible to crush, shear, and fray, it is natural to wonder if immunohistochemical and anatomical methods disturb or damage the cells they are designed to examine. Tissue fixation is typically incorporated to guard against this damage and is therefore critically important to the quality and significance of the harvested data. Here, we describe mechanisms of fixation; advantages and disadvantages of using formaldehyde and glutaraldehyde as fixatives during immunohistochemistry; and modifications of widely used protocols that have recently been found to improve cell shape preservation and immunostaining patterns, especially in proximal retinal neurons.
AB - Immunohistochemical and exvivo anatomical studies have provided many glimpses of the variety, distribution, and signaling components of vertebrate retinal neurons. The beauty of numerous images published to date, and the qualitative and quantitative information they provide, indicate that these approaches are fundamentally useful. However, obtaining these images entailed tissue handling and exposure to chemical solutions that differ from normal extracellular fluid in composition, temperature, and osmolarity. Because the differences are large enough to alter intercellular and intracellular signaling in neurons, and because retinae are susceptible to crush, shear, and fray, it is natural to wonder if immunohistochemical and anatomical methods disturb or damage the cells they are designed to examine. Tissue fixation is typically incorporated to guard against this damage and is therefore critically important to the quality and significance of the harvested data. Here, we describe mechanisms of fixation; advantages and disadvantages of using formaldehyde and glutaraldehyde as fixatives during immunohistochemistry; and modifications of widely used protocols that have recently been found to improve cell shape preservation and immunostaining patterns, especially in proximal retinal neurons.
KW - Antigen retrieval
KW - Fixatives
KW - Formaldehyde
KW - Glutaraldehyde
KW - PH
KW - Quenching
KW - Retina
KW - Sucrose
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UR - http://www.scopus.com/inward/citedby.url?scp=84939266578&partnerID=8YFLogxK
U2 - 10.1016/j.preteyeres.2015.04.001
DO - 10.1016/j.preteyeres.2015.04.001
M3 - Article
VL - 48
SP - 181
EP - 202
JO - Progress in Retinal and Eye Research
JF - Progress in Retinal and Eye Research
SN - 1350-9462
ER -