{"id":259,"date":"2012-04-10T16:48:14","date_gmt":"2012-04-10T16:48:14","guid":{"rendered":"https:\/\/wordpress.clarku.edu\/debrobertson\/?page_id=259"},"modified":"2012-05-23T17:18:40","modified_gmt":"2012-05-23T17:18:40","slug":"counting-cells-using-a-hemacytometer","status":"publish","type":"page","link":"https:\/\/wordpress.clarku.edu\/debrobertson\/laboratory-protocols\/counting-cells-using-a-hemacytometer\/","title":{"rendered":"Counting Cells Using a Hemacytometer"},"content":{"rendered":"

1.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Do not touch the face of the coverslip or the hemacytometer face with your fingers. Grease from you fingers can prevent the chamber from filling properly.<\/p>\n

 <\/p>\n

2.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Rinse the hemacytometer and coverslip with distilled water, dry using kimwipes and clean with lens paper.<\/p>\n

 <\/p>\n

3.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Mix sample with Pasteur pipette.\u00a0 With the coverslip in place, use a Pasteur pipette to transfer a small amount of the cell suspension to the counting chamber. Carefully touch the edge of the coverslip with the pipette tip and allow the chamber to fill by capillary action. Repeat with other chamber, mixing sample well before loading.<\/p>\n

 <\/p>\n

4.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Count cells in the four corner squares and the middle square (see diagram below)<\/p>\n

 <\/p>\n

5.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Each square of the hemacytometer (with cover slip in place) represents a volume of 0.1 mm3<\/sup>\u00a0or 10-4\u00a0<\/sup>cm3<\/sup>. Since 1 cm3<\/sup>\u00a0is equivalent to 1 mL, the subsequent cell concentration per mL (and the total number of cells) will be determined using the following calculations.<\/p>\n

 <\/p>\n

\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Cells per mL<\/em>= the average number of cells per square x 104<\/sup><\/p>\n

Example: If the average count per square is 45 cells, the density of cells is 45 x 104<\/sup>\u00a0cell mL-1<\/sup><\/p>\n

If the sample was diluted prior to counting, you should multiple the number above by the dilution factor<\/p>\n

Total cell number<\/em>\u00a0= cells per mL x the original volume of fluid from which cell sample was removed.<\/p>\n

Example: 4.5 x 105\u00a0<\/sup>cells mL-1<\/sup>\u00a0x 10 mL (culture volume) =4.5 x 106<\/sup>\u00a0cells total in the culture volume.<\/p>\n

 <\/p>\n

6.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Clean the hemacytometer with distilled water, dry, and place in storage box.<\/p>\n

 <\/p>\n

Reference:\u00a0Sigma Catalog (2000-2001) page 1849<\/p>\n

\"\"<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"

1.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Do not touch the face of the coverslip or the hemacytometer face with your fingers. Grease from you fingers can prevent the chamber from filling properly.   2.\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Rinse the hemacytometer and coverslip with distilled water, dry using kimwipes and clean …<\/p>\n

Counting Cells Using a Hemacytometer<\/span> Read More »<\/a><\/p>\n","protected":false},"author":72,"featured_media":0,"parent":8,"menu_order":70,"comment_status":"closed","ping_status":"closed","template":"onecolumn-page.php","meta":{"ngg_post_thumbnail":0,"footnotes":""},"class_list":["post-259","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages\/259","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/users\/72"}],"replies":[{"embeddable":true,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/comments?post=259"}],"version-history":[{"count":0,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages\/259\/revisions"}],"up":[{"embeddable":true,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages\/8"}],"wp:attachment":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/media?parent=259"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}