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The use of Lucifer Yellow and Mini-Ruby for intracellular staining in fixed brain tissue: methodological
1Zentrum der Morphologie, J.W. Goethe-Universität, Frankfurt am Main, Germany.
Abstract:
The quality of intracellular filling of Mini-Ruby (MR) is comparable to that of Lucifer Yellow (LY) in both perfusion-fixed rat and immersion-fixed autopsy human tissue. In adult human brain, the use of MR avoids the co-conversion of the typical intracellular lipofuscin deposits as is invariably the case during the photoconversion of LY. In autopsy studies (n = 35), the quality of intracellular staining does not correlate with age (range: 25-91 years) or the postmortem delay (range: 7-50 h). A short agonal status, however, is advantageous.
Insights
Mini-Ruby (MR) offers comparable intracellular filling quality to Lucifer Yellow (LY) in fixed tissues. MR is advantageous for adult human brain staining, preventing lipofuscin artifact seen with LY.
Area of Science:
- Neuroscience
- Histology
- Biochemistry
Background:
- Lucifer Yellow (LY) is a widely used fluorescent dye for intracellular filling.
- LY photoconversion can lead to artifactual co-conversion of lipofuscin deposits in human brain tissue.
- Alternative dyes are needed for reliable staining in specific applications.
Purpose of the Study:
- To compare the intracellular filling quality of Mini-Ruby (MR) with LY.
- To evaluate the utility of MR in adult human brain tissue, specifically regarding lipofuscin artifact.
- To assess the influence of age and postmortem delay on MR staining quality.
Main Methods:
- Perfusion fixation of rat tissue.
- Immersion fixation of autopsy human tissue.
- Microscopic evaluation of intracellular staining quality with MR and LY.
Main Results:
- Mini-Ruby (MR) demonstrated intracellular filling quality comparable to Lucifer Yellow (LY) in both rat and human tissues.
- In adult human brain, MR staining avoided the co-conversion of lipofuscin deposits, unlike LY.
- Staining quality with MR did not correlate with patient age or postmortem delay in autopsy studies (n=35).
- A short agonal status was found to be advantageous for staining quality.
Conclusions:
- Mini-Ruby is a viable alternative to Lucifer Yellow for intracellular filling in fixed tissues.
- MR offers a significant advantage for staining adult human brain tissue by preventing lipofuscin-associated artifacts.
- The efficacy of MR staining is robust across a range of ages and postmortem intervals, with optimal results in cases with short agonal periods.