Glass Slide Indexing System for making manual Microarrays using a VP 478 or a VP 478A Replicator

Glass Slide Indexing System for microarrays, uses 1″ x 3″ or 25mm x 75mm glass slides, compatible with standard FP1S6 pins

V&P Scientific is proud to introduce the first economical and practical way to make microarrays on glass slides for under $7,000.00 including accessories. Now you can make your own DNA or protein microarrays without buying a $20,000 machine. Whether you want to make large arrays on glass slides, work out new protocols for your robotic Glass Slide Arrayer or just Q/C new DNA preparations, this simple and flexible system allows you to tackle both large and small jobs without major setup hassles. Now you can have:

• Convenience
  Print your own microarrays anytime from 96 or 384 well plates
• Versatility
  Spot your samples as duplicates, triplicates or more
• Enhanced Features
  Print up to 768 spots per array. Print duplicate slides at the same time.
• Spot Size
  ~500 microns in diameter on silanated slides and 780 microns on Epoxy coated slides
• Spot Pitch
  1,125 microns on horizontal axis and 750 microns on the vertical axis, can change pitch
  easily for different spot sizes by skipping rows or columns
• Accessories Available
  Indexing System for 96 and 384 well source plates to simplify arraying and enhance spot size reproducibility. Replicator pin wash station to enhance arraying speed.
• Custom Options
  Can be adapted to your robotic system, change pins for different applications, e.g., larger volumes

Our Floating Pin Replicator can be fixtured with either 2 rows of 4 pins (8 total) on 9 mm centers for 96 well microplates (VP 478) see photo #1 or 4 rows of 8 pins (32 total) on 4.5 mm centers for 384 well microplates (VP 478A) see photo #2. When used in conjunction with the unique V&P Scientific Glass Slide Indexing System VP 470 (see photo #12 and photo #10) (US Patent #6,613,284 and Republic of China Invention Patent # NI-164,388, and European patent #1,363,737), this dual slide system places 768 spots ~500 microns in diameter onto a grid 18 mm wide and 36 mm tall on two separate slides. The ability to index two slides at the same time facilitates your ability to make duplicate slides. This also saves the time and effort required to wash the pins between slides. The pitch or center to center spacing between the spots is 750 microns on the Y axis and 1,125 microns in the X axis. This array will accommodate eight, 96 well microplates or two, 384 well microplates. However, you can use as few spots as your library requires or as spot size dictates. Our Replicator pins [FP1S6] have a 6 nl slot and liquid is transferred in that slot as well as in a hanging drop at the tip of the pin and on the sides of the pin [up to ~30 nl]. The diameter of the floating pins are 457 microns The pins must be reloaded after each spotting. Reloading the pins is as simple as dipping in a microplate. The amount that is deposited on the slide will depend on the surface tension of the liquid and the surface tension of the slide. Usually between 3 and 13 nl is on a slide. The volume deposited on a membrane will be larger due to the capillary action of the membrane. The spot produced on silanated slides is usually ~ 500 microns in diameter. This spot size allows a 24 by 32 array or 768 spots/array; while on epoxy coated slides, the spot is ~780 microns in diameter, so the highest density is 384 spots/epoxy coated slide.

The glass slide in photo #3 illustrates the 768 spotting pattern on a silanated glass slide made by Xenopore. This slide was photographed against a white background to better illustrate the spots.

The glass slide in photo #4 is an epoxy coated slide from Xenopore spotted with 5 mm 24mer oligonucleotide with a 3′ amine linker and a 5′ Cy5 label in 50% DMSO. GenePix 3 software was used to determine a mean spot diameter of 780 microns with a CV of 5.5%. Photo #4 is an enlargement of the slilde in photo #5.

Accuracy and Precision Data of Spot Diameter from the Slide in photo #5

Position Mean SD %CV Max Mi5

_____ _____ ____ _____ ____ ___

Pin 1 793.1 42.7 5.4% 920 700

Pin 2 786.7 42.1 5.4% 910 700

Pin 3 789.2 40.5 5.1% 890 710

Pin 4 786.3 35.4 4.5% 870 730

Pin 5 787.5 39.1 5.0% 910 710

Pin 6 757.9 41.4 5.5% 870 690

Pin 7 795.8 41.7 5.2% 930 730

Pin 8 745.2 37.4 5.0% 840 690

TOTAL 780.2 43.3 5.5% 930 690

Photo #6 is an Aldehyde (Silanated) Slide with Amino/CY5-Labeled Oligo 768 spot array, 5′ Cy5 and 3’Amino linker.

We have also obtained data using SigmaScreen Silanated slides with a 24mer 5uM Oligo with a 5′-Cy5 label spotted in 50% Array It and 50% 15 mM Sodium Phosphate buffer and scanned on an Applied Precision (API) Array WoRx Scanner. BioDiscovery software was used to determine a mean signal area of 7,914 with a CV of 5.7% and a mean signal of 40,434 with a CV of 4.3%.

One of our customers uses the Glass Slide Arrayer to make Protein arrays, Tumor Tissue arrays and Tumor mRNA arrays.

Microarrays on Membrane Slides

The VP 470 Glass Slide Indexer has a unique spring loaded locating feature (see photo #7) that will accommodate either 1″ x 3″ or 25 mm x 75 mm glass slides1 mm thick. The spring loaded locator will allow irregular slides up to 25.9 mm (1.02″) wide and 76.36 mm (3.007″) long to be located. Thinner or thicker slides can also be accommodated. The diameter of the floating pins are 457 microns The pins must be reloaded after each spotting. Reloading the pins is as simple as dipping in a microplate.

If you are transferring DNA, cDNA or other materials in solutions with high surface tension, you will find that our new V&P Pin Cleaner treatment of the pins will greatly increase the precision of delivery. For an explanation, see the FAQ page and the V&P Pin Cleaner page.

Complete technical notes are available for the Glass Slide Microarrayer

Photo #9 illustrates the 8 place horizontal X axis indexing holes and the 12 place vertical Y axis indexing holes with their respective pins. Changing an index position is as simple as moving the pin to the next hole. This ingenious indexing system is capable of exacting precision and accuracy. Our unique indexing system also guards against the accidental misplacement of the Indexing pin into the wrong indexing hole. The two sets of guide pins are used to locate the VP 478 and VP 478A Arrayers onto the two glass slide bays of the VP 470 Glass Slide Indexer. The guide pins locate the Arrayers by fitting into holes in the bottom of the Arrayers. This provides a very rapid yet simple and easy way to locate the Arrayers. Another “fool proof” system incorporated into the design is that the pair of guide pins have different diameters thus preventing the Arrayer from being placed in the slide bay in the reversed position. (US Patent #6,613,284, Republic of China Invention Patent # NI-164388, European patent #1,363,737).

Photo #10 demonstrates how the moving of index pins creates a high density array.

The VP 470 Glass Slide Indexer is very compact measuring only 15 cm width, 13 cm in depth and 4 cm tall. It is constructed of anodized aluminum with stainless steel pins and weighs 1.078 Kilo grams. The VP 478 and VP 478R Glass Slide Replicators measure 8.4 cm in width, 2 cm in depth and 11 cm tall, see photo #11. They are constructed of delrin with stainless steel pins. The VP 478 weighs only 167 grams and the VP 478A weighs 230 grams. The VP 478A and VP 478AR Glass Slide Replicators measure 8.4 cm in width, 2.5 cm in depth and 11 cm tall. They are also constructed of delrin with stainless steel pins and weighs 230 grams. Both units can be disinfected by bleach and alcohol treatment.

Because of the unique way the pins are positioned in the two Glass Slide Replicators 8 pin – VP 478 -9 mm pitch, 96 well, and 32 pin – VP 478A – 4.5 mm pitch, 384 well, they each have to be used with their specific Microplate Indexing System.

Glass Slide Replicators with longer pins to reach the bottom of some tall PCR plates are now available as well as a specially adapted Glass Slide Indexer for accommodating the longer pins. Just order the VP 478T with longer FP1S6T pins see photo #13.

The Standard Glass Slide Index can be modified with longer alignment pins and extension blocks to adjust the proper distance from the pins to the glass slide. This version for the taller pins used with deep well microplates is the VP 470T, see photo #14.

Photo #13 illustrates one of several custom modifications that can be done with the VP 478 or VP 478A. Larger or smaller diameter spots can be made by simply changing the diameter of the pin. We have done many other custom modifications using larger [1.58 mm] diameter pins and pins as small as 0.229 mm diameter.

Accessories for the Manual Glass Slide Microarrayer

Microplate Indexing System
We provide for your convenience a very simple method of registering the Replicator pins to the center of the microplate wells see photo #8. This feature is key in making consistent diameter spots. The microplate indexing system also helps you keep track of your position in the source plate.

Replicator Pin Wash and Blot Station
We provide for your convenience an efficient device to wash and clean the Replicator pins between spotting.

Cleaning Pins

We strongly recommend that you use a lint free blotting paper such as VP 522 or VP 522A to clean the Replicator pins between spotting operations. The presence of lint on the pins will affect the spot size on the slide. We also recommend that you use the VP 110 Pin Cleaning Solution to periodically clean the Replicator pins.

Coming Soon

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