公开(公告)号：US20140127744A1

公开(公告)日：2014-05-08

申请号：US14154766

申请日：2014-01-14

Applicant: Essen Instruments, Inc.

Inventor： Kirk Schroeder ,  Bradley D. Neagle

IPC: G01N33/50

CPC classification number: C12Q1/02 ,  B01L3/50255 ,  B01L3/5085 ,  B01L2200/0647 ,  B01L2200/142 ,  B01L2300/0654 ,  B01L2300/0681 ,  B01L2300/0829 ,  B01L2300/163 ,  G01N21/6452 ,  G01N21/6458 ,  G01N33/5008 ,  G01N33/5029 ,  G01N33/54366 ,  G01N33/58

Abstract: Apparatus and methods to improve the Boyden chamber used in cellular biological measurements, allowing quantitative optical microscopy of biological cells in situ without using fluorescent probes or optical staining. In the preferred embodiment, a thin porous membrane separating top and bottom reservoirs includes an array of precisely positioned micropores pores manufactured using a laser-based photo-machining (ablation) process. The membrane may be composed of polyethylene terephthalate (PET), polycarbonate, polyimide, polyether ether ketone (PEEK) or other appropriate material. The pores formed in the membrane may have diameters in the range of 1 to 15 microns and spaced apart at a distance ranging from 10 to 200 microns. A plurality of upper and lower reservoirs may be provided to form a multi-well plate. The invention finds application in a wide range of potential biological applications where Boyden chamber geometries are currently used including co-culture studies, tissue remodeling studies, cell polarity determinations, endocrine signaling, cell transport, cell permeability, cell invasion and chemotaxis assays.

Abstract translation: 用于改善细胞生物学测量中使用的Boyden室的装置和方法，允许原位生物细胞的定量光学显微镜检查，而不使用荧光探针或光学染色。 在优选实施例中，分离顶部和底部储存器的薄多孔膜包括使用基于激光的光加工（消融）工艺制造的精确定位的微孔的阵列。 膜可以由聚对苯二甲酸乙二醇酯（PET），聚碳酸酯，聚酰亚胺，聚醚醚酮（PEEK）或其它合适的材料组成。 形成在膜中的孔可以具有在1至15微米范围内的直径并且间隔开10至200微米的距离。 可以设置多个上部和下部储存器以形成多孔板。 本发明应用于广泛的潜在生物应用，其中目前使用Boyden室几何形状，包括共培养研究，组织重塑研究，细胞极性测定，内分泌信号，细胞转运，细胞通透性，细胞侵袭和趋化性测定。