A useful low temperature method for post-embedding electron immunocytochemistry in routine histopathology

1. A modification of the digitonin method of Zuurendonk & Tager (1974) (Biochim. Biophys. Acta 333, 393-399) (i.e. the ‘convaentional’ method) was developed that allows the fractionation of isolated hepatocytes at −5 degrees C (i.e. ‘low-temperature’ method). 2. With respect to compartmentation of adenine nucleotides, glutamate and citrate, the two methods yielded very similar results. 3. In contrast, the mitochondrial amounts of aspartate and malate, as revealed by the low-temperature method, were about twice as high as those found by the conventional procedure. No change in the total cellular content occurred. 4. With n-butylmalonate and glisoxepid present in the conventional digitonin medium, significantly higher amounts of malate and aspartate respectively were found in the mitochondrial pellets. The results obtained by the low-temperature method, however, were not influenced by the these inhibitors. 5. It is concluded that under the conventional conditions of cell fractionation no appreciable redistribution of adenine nucleotides, glutamate and citrate occurs.

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RETRACTED: Low temperature method for synthesis of ZnS quantum dots and its luminescence characterization studies

Applied Surface Science ◽

10.1016/j.apsusc.2012.09.026 ◽

2013 ◽

Vol 264 ◽

pp. 17-20 ◽

Cited By ~ 8

Author(s):

K. Senthilkumar ◽

T. Kalaivani ◽

S. Kanagesan ◽

V. Balasubramanian

Keyword(s):

Quantum Dots ◽

Low Temperature ◽

Temperature Method ◽

Characterization Studies

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Micro-morphological Characterization of Fresh Cannabis sativa L. Roots After Lead Exposure Using Low Temperature Method for ESEM

Microscopy and Microanalysis ◽

10.1017/s1431927618007018 ◽

2018 ◽

Vol 24 (S1) ◽

pp. 1306-1307

Author(s):

Vilem Nedela ◽

Eva Tihlafikova ◽

Biljana Dordevic ◽

Nikola Slamova

Keyword(s):

Low Temperature ◽

Lead Exposure ◽

Cannabis Sativa ◽

Morphological Characterization ◽

Temperature Method

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A low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis

Biochemical Journal ◽

10.1042/bj2800331 ◽

1991 ◽

Vol 280 (2) ◽

pp. 331-334 ◽

Cited By ~ 76

Author(s):

N Flint ◽

F L Cove ◽

G S Evans

Keyword(s):

Small Intestine ◽

Low Temperature ◽

Intestinal Epithelium ◽

Sequential Fractionation ◽

Potential Value ◽

Temperature Method ◽

Small Intestinal ◽

The Comparative Study ◽

Cell Phenotypes ◽

Proliferative Cell

A variety of enzymic and non-enzymic methods to isolate epithelium from the small intestine have been previously published. Sequential fractionation of cells from the villus to the crypt has been reported in some of these papers, which allows the comparative study of terminally differentiated and proliferative cell phenotypes. However, these methods often involve the incubation of tissues at 37 degrees C, which may affect the structural and biochemical integrity of the cells. We have developed a rapid low-temperature (4 degrees C) method for isolating purified populations of crypt and villus cells from mouse and rat intestines. The fractionated cells have been partially characterized, and the potential value of the procedure has been indicated by the ability to analyse the comparative protein and mRNA expression along the crypt-villus axis.

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