Pagliaro L, Reitz F, Wang J
Center for Bioengineering, WD-12 University of Washington, Seattle, USA.
Undersea Hyperb Med. 1995 Jun;22(2):171-81.
We have developed an optical pressure chamber designed for use with high numerical aperture oil immersion microscope objectives at working pressures up to 1,000 psi (67 atm abs). The chamber is optimized for studies of living, adherent, cultured mammalian cells using high resolution epifluorescence and phase contrast microscopy, and biophysical techniques such as fluorescence redistribution after photobleaching and optical trapping. The primary optical window assembly of the chamber can be removed and placed into a standard 35-mm tissue culture dish, allowing for culture, microinjection, and micromanipulation of adherent cells before they are loaded into the chamber. The chamber is designed to fit into a commercially available stage heater for temperature control, and we used a computer-controlled high pressure liquid chromatography pump for pressure control. A graphic software interface allows the user to program "dive" profiles and to link temperature and pressure data with digital image files of specimens under study. A minor modification of the present design will allow perfusion at high pressure.
我们开发了一种光学压力室，设计用于与高数值孔径油浸显微镜物镜配合使用，工作压力高达1000 psi（67 atm绝对压力）。该压力室经过优化，适用于使用高分辨率落射荧光和相差显微镜以及生物物理技术（如光漂白后的荧光重新分布和光镊）对活的、贴壁的、培养的哺乳动物细胞进行研究。压力室的主要光学窗口组件可以拆卸并放入标准的35毫米组织培养皿中，以便在将贴壁细胞加载到压力室之前对其进行培养、显微注射和显微操作。压力室设计为可安装到市售的载物台加热器中进行温度控制，我们使用计算机控制的高压液相色谱泵进行压力控制。图形软件界面允许用户编程“潜水”曲线，并将温度和压力数据与所研究标本的数字图像文件链接起来。对当前设计进行 minor 修改将允许在高压下进行灌注。
