A simple one-step procedure for freezing tissue for light microscopy

L. Terracio; H. D. Coulter

A simple one-step procedure for freezing tissue for light microscopy

L. Terracio, H. D. Coulter

Molecular Pathobiology

Research output: Contribution to journal › Article › peer-review

Original language	English (US)
Pages (from-to)	45-47
Number of pages	3
Journal	Stain Technology
Volume	53
Issue number	1
State	Published - 1978

ASJC Scopus subject areas

Anatomy

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@article{fe0ece9b604447ae908704598ae88e70,

title = "A simple one-step procedure for freezing tissue for light microscopy",

author = "L. Terracio and Coulter, {H. D.}",

note = "Funding Information: The support of this work by the American Cancer Society under Grant No. PDT-42A is gratefully acknowledged. Funding Information: We previously reported (Cryobiology 12, 584 ( 1975) ) on the separation of granulocytes from whole human blood using the Elutriator rotor (Beckman Company). We have further improved and standardized this method and also introduced a new way of collecting and separating blood cells without the use of anticoagulants. To prevent clotting, freshly collected blood is immediately diluted lo-fold with Hanks{\textquoteright} balanced salt solution. The diIuted blood is then loaded directly into the elutriation chamber without the use of a mixing chamber. Only two flow rates are required for the separation of granulocytes: the priming flow rate, ranging from 11 to 14 ml, and the elutriating flow rate, ranging from 14 to 35 ml. Because of the variations in individual blood samples, visual monitoring of the cell distribution within the chamber is required for the precise adjustment of the flow rate to achieve the maximal cell separation and cell yield. Purified granulocytes are eluted with minimal dilution by reversing the direction of the flow through the separation chamber rather than by flushing out the cells with an increased flow rate. The whole procedure of separating granulocytes from 5 ml of bbod takes about 25 min. Yields of pure granulocytes as high as 100% are obtainable. Granulocytes isolated by the elutriation procedure at a rotor speed of 2500 rpm are indistinguishable from dextran-sedimented cells in terms of membrane integrity, phagocytic activity, and bactericidal function. These granulocytes also do not clump as readily as the dextran-sedimented cells. Higher rotor speeds, up to 5000 rpm, during elutriation decreases the bactericidal{\textquoteright} ability of granulocytes. We believe this method offers a new and effective tool for studying granulocyte function by providing cells in concentrated suspension with minimal handling. Supported by NIH Grant ROl CA12754-05.",

year = "1978",

language = "English (US)",

volume = "53",

pages = "45--47",

journal = "Stain Technology",

issn = "0038-9153",

publisher = "Informa Healthcare",

number = "1",

}

TY - JOUR

T1 - A simple one-step procedure for freezing tissue for light microscopy

AU - Terracio, L.

AU - Coulter, H. D.

N1 - Funding Information: The support of this work by the American Cancer Society under Grant No. PDT-42A is gratefully acknowledged. Funding Information: We previously reported (Cryobiology 12, 584 ( 1975) ) on the separation of granulocytes from whole human blood using the Elutriator rotor (Beckman Company). We have further improved and standardized this method and also introduced a new way of collecting and separating blood cells without the use of anticoagulants. To prevent clotting, freshly collected blood is immediately diluted lo-fold with Hanks’ balanced salt solution. The diIuted blood is then loaded directly into the elutriation chamber without the use of a mixing chamber. Only two flow rates are required for the separation of granulocytes: the priming flow rate, ranging from 11 to 14 ml, and the elutriating flow rate, ranging from 14 to 35 ml. Because of the variations in individual blood samples, visual monitoring of the cell distribution within the chamber is required for the precise adjustment of the flow rate to achieve the maximal cell separation and cell yield. Purified granulocytes are eluted with minimal dilution by reversing the direction of the flow through the separation chamber rather than by flushing out the cells with an increased flow rate. The whole procedure of separating granulocytes from 5 ml of bbod takes about 25 min. Yields of pure granulocytes as high as 100% are obtainable. Granulocytes isolated by the elutriation procedure at a rotor speed of 2500 rpm are indistinguishable from dextran-sedimented cells in terms of membrane integrity, phagocytic activity, and bactericidal function. These granulocytes also do not clump as readily as the dextran-sedimented cells. Higher rotor speeds, up to 5000 rpm, during elutriation decreases the bactericidal’ ability of granulocytes. We believe this method offers a new and effective tool for studying granulocyte function by providing cells in concentrated suspension with minimal handling. Supported by NIH Grant ROl CA12754-05.

PY - 1978

Y1 - 1978

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M3 - Article

C2 - 663947

AN - SCOPUS:0017897466

SN - 0038-9153

VL - 53

SP - 45

EP - 47

JO - Stain Technology

JF - Stain Technology

IS - 1

ER -

A simple one-step procedure for freezing tissue for light microscopy

ASJC Scopus subject areas

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