Bacteria Counting Chamber is a one piece construction ensuring durability and accuracy with a Thoma ruling on a single round plateau. The Bacteria Counting Chamber is used for bacteria and sperm counting. Cell Depth is 0.02mm. Usage Bacteria/Sperm Counts Cell Depth 0.02mm +/- 1% (1/50mm) Volume of 1 Square mm 0.02 Microliter Ruling Pattern Thoma, 1/400 Square mm Rhodium coated (bright-line) No Counting Technique : Thoma Counting assumes precise knowledge of the limit lines of the counting chambers used. These are shown in the illustration below. Each one of the small squares is 0.05 x 0.05 mm square, ie 1/400 mm2. This figure will be found printed onto the surface of the counting chamber. To ensure that cells which are on or along the limit lines are not counted twice or are not missed during the count, certain rules have to be observed (see illustration below – this illustration is for an Improved Neubauer, hence the triple lines, but the principle is the same for the Thoma).

The black cells are the ones which would be included in the count for that square, the white ones would not as they will be included in the count for the adjacent squares. The count should be started at the top left-hand corner and follow the direction shown by the arrow (see illustration). Counting may be enhanced with the microscopes illumination reduced.

Notes on Counting Use reduced microscope illumination for all chambers.
The difference of the counter cells in the large squares and the group squares must not exceed 10 cells. Double checks must be performed for all cell counts. After
counting the two counting nets the bottom counting net is to be counted in the same way as a check. When doing this it is to be ensured that the chamber has not dried out. This can be prevented by filling the bottom chamber only shortly before the count and not counting after the sedimentation time.
   The difference between the totals of the counts for the two counting nets must not exceed 10 cells. The average value of the counts is then used in the calculation formula or multiplied by the corresponding factor.

計(jì)數(shù)板的使用

準(zhǔn)備
準(zhǔn)備計(jì)數(shù)時(shí)，磨光的表面小心地用擦鏡紙擦干凈。蓋玻片也要擦干凈。

蓋上蓋玻片在需要蓋的周邊區(qū)域需要用蒸餾水弄的微濕然后蓋玻片要慢慢的從計(jì)數(shù)板的前端推入，直到蓋玻片的前端邊緣與計(jì)數(shù)板的劃格子的區(qū)域處于同位。

注意：蓋玻片是很容易壞掉的！

在外部支持和蓋玻片之間的干擾線（牛頓環(huán)）的組成顯示了蓋玻片的準(zhǔn)確位置。

計(jì)數(shù)板加樣

取一個(gè)良好混合型的吸管并且放置一些初期的幾滴液體。

把吸管的外部擦干然后保持吸管一定的角度直到小滴液體被吸管吸上去。然后這滴液體被放置在蓋玻片和計(jì)數(shù)板之間。由于蓋玻片和計(jì)數(shù)板之間的毛細(xì)管現(xiàn)象作用的結(jié)果，兩個(gè)平面之間的縫隙被灌滿。在溶液要滿出擊數(shù)板截面的邊緣之前，馬上移走吸管的。如果有可見氣泡或者液體已經(jīng)滿出邊緣并且進(jìn)入其他的溝里，那么計(jì)數(shù)板就要擦干凈并且重新加樣。

計(jì)數(shù)

裝載好的計(jì)數(shù)板被放置在顯微鏡臺(tái)上然后計(jì)數(shù)格在低倍鏡焦距中顯示。高倍物鏡要非常小心的操作，因?yàn)橛?jì)數(shù)板比常規(guī)的要更厚。如果你不小心的話，會(huì)導(dǎo)致計(jì)數(shù)板或者物鏡鏡頭的損傷。

如果可能的話，在作血細(xì)胞計(jì)數(shù)的時(shí)候使用機(jī)械平臺(tái)的顯微鏡。顯微鏡的照明是重要的，太亮了會(huì)影響劃格線的觀察。光可以通過虹膜光圈減弱直到劃得格子在背景中再次清晰可見。細(xì)胞/質(zhì)粒的計(jì)數(shù)應(yīng)該充分稀釋到他們不再互相重疊，并且均勻的分配。

要完成計(jì)數(shù)檢測(cè)需要擴(kuò)大到期望的可見的細(xì)胞/質(zhì)粒。細(xì)胞數(shù)量的計(jì)算推薦計(jì)數(shù)100方格作為排除少量的結(jié)果。

計(jì)數(shù)格在超過劃線區(qū)域使用時(shí)需要均勻的分配，并且這個(gè)通過藉由三倍之?dāng)?shù)決定的方格支架在進(jìn)入?yún)^(qū)段之內(nèi)。

計(jì)數(shù)完100方格后總數(shù)除100（計(jì)數(shù)的方格數(shù)），來得到每格的平均值。乘稀釋度，除1/4000（每個(gè)小方格的立方容積1/20 x 1/20 x 1/10 – 1/4000mm3）。這次計(jì)算得到的數(shù)字是原非稀釋流質(zhì)的每立方毫米的細(xì)胞數(shù)量。