Hausman G J
Stain Technol. 1981 May;56(3):149-54. doi: 10.3109/10520298109067302.
Various fixatives as well as tissue and slide handling procedures have been evaluated in attempts to demonstrate adipocytes histochemically while maintaining cell and tissue integrity. The optimal procedure for analysis of immature adipose depots consists of the following steps: 1) fresh, unfixed tissues are rapidly in isopentane quenched in a liquid nitrogen bath; 2) cryostat sections are cut, removed from the knife with a room temperature slide, and then air dried for 5-10 minutes; 3) slides can be stained directly with picro-Ponceau or toluidine blue procedures or with oil red O following fixation for 30 minutes in cold (4 C) 10% formalin-CaCl2 (1.25%). For analysis of mature rat adipose depots steps 2 and 3 are modified as follows: 2) cryostat sections are removed from the knife with a cold slide (-20 C) and dried for 30 minutes at 4 C; 3) the mounted sections are stained with oil red O following fixation for 30 minutes in cold (4 C) 10% formalin-HgCl2 (2.5%). When procedures described above for immature adipose depots are combined with esterase staining, adipocyte cytoplasm is clearly demonstrated. These procedures allow the routine use of fresh frozen, unfixed cryostat sections in studies of adipose cellularity.
为了在组织化学上显示脂肪细胞，同时保持细胞和组织的完整性，人们对各种固定剂以及组织和玻片处理程序进行了评估。分析未成熟脂肪库的最佳程序包括以下步骤：1）将新鲜的、未固定的组织迅速放入异戊烷中，在液氮浴中骤冷；2）用冷冻切片机切片，用室温玻片从切片刀上取下，然后空气干燥5-10分钟；3）玻片可以直接用苦味酸-丽春红或甲苯胺蓝程序染色，或者在冷（4℃）的10%福尔马林-CaCl₂（1.25%）中固定30分钟后用油红O染色。对于成熟大鼠脂肪库的分析，步骤2和3修改如下：2）用冷玻片（-20℃）从切片刀上取下冷冻切片机切片，并在4℃下干燥30分钟；3）将裱贴好的切片在冷（4℃）的10%福尔马林-HgCl₂（2.5%）中固定30分钟后，用油红O染色。当上述未成熟脂肪库的程序与酯酶染色相结合时，脂肪细胞质可清晰显示。这些程序允许在脂肪细胞研究中常规使用新鲜冷冻的、未固定的冷冻切片机切片。
