Apparatus for the handling of electrophoretic transfer membranes and for the chemiluminescent detection of blots contained thereon

• US 5,306,468 A
• Filed: 07/19/1991
• Issued: 04/26/1994
• Est. Priority Date: 07/19/1991
• Status: Expired due to Term

• Current Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Polsky, Shay E., Anderson, Lynne E., Kobos, Robert K., Robertson, Charles W.
• Primary Examiner(s): Johnston, Jill A.
• Application Number:
  US07/739,123
• Time in Patent Office: 1,012 Days
• Field of Search: 422/99, 422/101, 422/104, 204/182.8, 100/93 P
• US Class Current: 204/612
• CPC Class Codes: G01N 27/44717 Arrangements for investigating the separated zones, e.g. localising zones
• CPC Class Codes: G01N 27/44721 by optical means

Abstract

Improved apparatus for the handling and detecting of an electrophoretic transfer membrane is disclosed herein, the improvement comprising guide apparatus designed to accommodate the transfer membrane, a pressure head positioned relative to the guide apparatus to releasably engage the membrane, and light shielding apparatus positioned relative to the guide apparatus to selectively enclose about the transfer membrane. The apparatus is operated so that the pressure head engages a flexible portion of the transfer membrane containing an electrophoretic blot while it is in a position appropriate for detection. There is also disclosed herein a process for utilizing the apparatus of the invention. In the process drying and optionally heating of the membrane where the blot has been contacted with enzyme-triggered 1,2-dioxetanes, to enhance the resulting chemiluminescent signal, is disclosed.

Claims

1. In an improved apparatus for the handling of an electrophoretic transfer membrane and the detecting of a signal generated by separated biological materials on said electrophoretic transfer membrane that are suitably treated to generate said signal, wherein the electrophoretic transfer membrane comprises a rigid frame (i) that supports a flexible membrane (ii) along portions of the periphery thereof, the membrane (ii) containing thereon a detectable, signal generating electrophoretic blot, and a signal detector therefor, the improvement comprising:

(a) guide apparatus designed to accommodate the rigid frame (i) and the flexible membrane (ii) so that the flexible membrane (ii) is positioned for reading in a location for detection while engaged by pressure head (b);

(b) pressure head positioned with respect to said guide apparatus and said location to releasably engage the flexible membrane (ii);

(c) light shielding apparatus positioned with respect to said guide apparatus to selectively enclose about the rigid frame (i) and the flexible membrane (ii); and

(d) means for sequentially operating said guide apparatus (a), said pressure head (b), and said light shielding apparatus (c) so that the rigid frame (i) and the flexible membrane (ii) are enclosed and said pressure head engages the flexible membrane (ii) while the transfer membrane is in said location appropriate for detection.

2. The apparatus of claim 1 wherein said guide apparatus (a) further comprises a frame guide having a through slot aligned to receive the rigid frame (i) and the flexible membrane (ii) for engagement of said flexible membrane (ii) by said pressure head (b) and to discharge the rigid frame (i) and the flexible membrane (ii) after detection.

3. The apparatus of claim 1 wherein said pressure head is pivotally positioned with respect to said guide apparatus to releasably engage the flexible membrane (ii), via a moment arm rotatably secured to the apparatus.

4. The apparatus of claim 3 wherein said pressure head further comprises a pressure plate which contacts the flexible membrane (ii) and is shaped to bias the flexible membrane (ii) away from the rigid frame (i) during engagement, and optionally heating means secured to said pressure plate, said pressure plate and optional heating means being secured to said moment arm.

5. The apparatus of claim 2 wherein said through slot has first and second ends with said location for detection therebetween and said first end is in a region of reception of the rigid frame (i) and the flexible membrane (ii) and said second end is in a region of discharge of the rigid frame (i) and the flexible membrane (ii) and said light shielding apparatus comprises a first light shielding means located proximate to said first end of said through slot and having open and close positions, and a second light shielding means located proximate to said second end of said through slot and having open and close positions, operably connected so that as the rigid frame (i) and the flexible membrane (ii) are received within said through slot said first light shielding means is open and said second light shielding means is closed sufficient to stop the rigid frame (i) and the flexible membrane (ii) from passing thereby, during detection said first and second light shielding means are closed sufficient to shield light for detection, and as the rigid frame (i) and the flexible membrane (ii) are discharged from said through slot, said second light shielding means is open.

6. The apparatus of claim 5 wherein said first light shielding means comprises a first arm with a first blade at one end thereof in light shielding relationship to said through slot, and said second light shielding means comprises a second arm with a second blade at one end thereof in light shielding relationship to said through slot, and said first and second arms being operably connected at each other end thereof.

7. The apparatus of claim 6 wherein said guide apparatus (a) further comprises a cover having opposed entry and exit slots which accommodate the rigid frame (i) and the flexible membrane (ii), said entry slot being located proximate to said first end of said through slot with said first light shielding means interposed therebetween and said exit slot being located proximate to said second end of said through slot with said second light shielding means interposed therebetween.

8. In an improved apparatus for the handling and detecting of an electrophoretic transfer membrane that comprises a rigid frame (i) that supports a flexible membrane (ii) along portions of the periphery thereof, and the membrane (ii) containing thereon a detectable electrophoretic blot, the improvement comprising in combination:

(a) guide apparatus designed to accommodate the rigid frame (i) and the flexible membrane (ii) so that the flexible membrane (ii) is positioned for reading in a location for detection while engaged by pressure head (b);

(b) pressure head positioned with respect to said guide apparatus and said location to releasably engage the flexible membrane (ii);

(c) light shielding apparatus positioned with respect to said guide apparatus to selectively enclose about the rigid frame (i) and the flexible membrane (ii);

(d) means for sequentially operating said guide apparatus (a), said pressure head (b), and said light shielding apparatus (c) so that the rigid frame (i) and the flexible membrane (ii) are enclosed and said pressure head engages the flexible membrane (ii) while the transfer membrane is in said location appropriate for detection; and

(e) means for detecting the signal generated by the electrophoretic blot comprising a CCD camera focused upon the flexible membrane (ii) in said location while the flexible membrane (ii) is engaged by said pressure head, and with respect to said guide apparatus so that a suitable image of the signal generated by the blot is obtained during engagement of the flexible membrane (ii) by the pressure means.

9. The apparatus of claim 8 wherein a light excluding tube is interposed between said CCD camera and said guide apparatus (a) to establish focusing of a suitable image of the signal generated by the blot.