Bell G A, Mellor J D
Neurosci Lett. 1984 May 4;46(2):197-201. doi: 10.1016/0304-3940(84)90441-5.
The condition of freeze-fixed mitral cells from the main olfactory bulb of the rat brain was found to depend on the rat at which the tissue is frozen: at very slow rates, mitral cells swell and rupture; at faster rates, cell integrity is preserved. Cooling the tissue to just above its freezing point before rapid freeze-fixation helps to maintain cell integrity at depths well below the surface. When freeze-fixation must be used, as in the 2-deoxyglucose technique, rapid freezing rates should be used to minimize diffusion of isotope and increase autoradiographic resolution. After rapid freezing, better cell integrity was produced by freeze-drying and plastic embedding than by cryostat sectioning and thaw-mounting.
研究发现，大鼠脑主嗅球中经冷冻固定的二尖瓣细胞的状态取决于组织冷冻时的速率：在极慢的速率下，二尖瓣细胞会肿胀并破裂；在较快的速率下，细胞完整性得以保留。在快速冷冻固定之前将组织冷却至略高于其冰点有助于在远低于表面的深度维持细胞完整性。当必须使用冷冻固定时，如在2-脱氧葡萄糖技术中，应采用快速冷冻速率以尽量减少同位素扩散并提高放射自显影分辨率。快速冷冻后，冷冻干燥和塑料包埋比低温恒温器切片和解冻固定能产生更好的细胞完整性。
