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As
the trend towards assay miniaturization continues with increasing
demands to make nanoliter transfers, here is a simple way to convert a High
Resolution Engineering Robot into a 96, 384 or 1,536
pin tool.
The Protedyne Corporation
Robot delivers an entirely
new screening platform to address assays. These features enable users to save time and money, while addressing their
preferred screening targets.
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Procedures
for transferring liquids, robot
speed, wash solutions, blotting protocols
and pin cleaning can found at this
hyperlink.
Cleaning the pins between mother plates is easy and
effective. In
an experiment to determine the
amount of carry over after 100 transfers of concentrated FITC (mother
to daughter plate) we were only able to detect 0.000,001,8% of
the residual FITC on
the pin using only two static 100 ml wash baths. To put this
into
perspective, it is equivalent to removing all but 18 molecules out of a
possible 1,000,000,000 molecules. This experiment was
repeated
using DMSO as
the first wash solution and virtually identical results were obtained,
indicating the thoroughness of the wash.
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The
list on the right summarizes the factors
that contribute to the volume delivered. With each
application
these factors can be controlled and standardized so that the delivery
volumes are very reproducible. With most applications the
CV's
are less than 5%.
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V&P
pins transfer liquid by carrying the
liquid on the sides, tips and in slots we cut into the tips. The amount
delivered is determined by the diameter and surface tension of the pin,
the surface tension of the liquid, how far the pin is submerged into
the liquid and the speed with which the pin is removed from the
liquid. When all of these variables are fixed, the accuracy
of
the liquid transfer is very consistent, with CV averages better than 5%.
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Increasing
the speed of withdrawal from the
source plate by 7- fold will increase the volume delivered by as much
as 3 fold in a linear relationship. This phenomenon can be
exploited to expand the range of delivery volume for a single pin. The
table and the chart below illustrates the effects of increasing the
speed 7- fold. The speed range is from 0.78 cm/sec to 5.7
cm/sec. Use this chart to select the pin that will deliver in
your desired range.
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PIN
|
DIAMETER
|
SHAPE |
DELIVERY
VOLUME RANGE*# |
|
FP1 |
0.457
mm |
solid |
19
– 68 nl |
|
FP1S6 |
0.457 mm |
6
nl slot |
26
– 74 nl |
|
FP1S10 |
0.457
mm |
10
nl slot |
31
– 79 nl |
|
FP1S20 |
0.457
mm |
20
nl slot |
43
– 89 nl |
|
FP1S30 |
0.457
mm |
30
nl slot |
53
– 99 nl |
|
FP1S40 |
0.457
mm |
40
nl slot |
64
– 109 nl |
|
FP1S50 |
0.457
mm |
50
nl slot |
74
– 117 nl |
|
FP3 |
0.787
mm |
solid |
84
– 240 nl |
| FP3S100 |
0.787
mm |
100
nl slot |
178 –
326 nl |
|
FP3S200 |
0.787
mm |
200
nl slot |
278 – 424
nl |
|
FP3S500 |
0.787
mm |
500
nl slot |
527 – 656
nl |
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*#
delivery volume range is determined by speed of
withdrawal from source liquid.
slow speed = 0.78 cm/sec = low volume delivery range
fast speed = 5.70 cm/sec = high volume delivery
range
Follow this link to assay methods
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The
links below are to delivery volume range
tables for uncoated and hydrophobic coated delivering either DMSO or
Aqueous solutions.
DMSO
TRANSFERS
WITH HYDROPHOBIC COATED PINS
DMSO
TRANSFERS WITH
UNCOATED (STAINLESS STEEL) PINS
AQUEOUS
TRANSFERS WITH HYDROPHOBIC
COATED PINS
AQUEOUS
TRANSFERS WITH UNCOATED
(STAINLESS STEEL) PINS
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1.
What volume do you need to
transfer?
DMSO?
Aqueous?
See these hyperlinked tables to determine your options. Also
consider custom slot pins.
2. Will the source plate have wells with
significantly different levels of liquid? (Cherry picked
source
plate or edge drying effect?). If yes and if the absolute
volume
transferred is critical, then select the largest slot pin that is in
your transfer range. This will minimize the effect of liquid
height on the volume of liquid carried on the sides of the
pin.
Also consider custom slot pins.
3.
Does the material transferred bind
non-specifically to stainless steel? If yes then select the
Hydrophobic/lipophobic
coated pins. If no,
select the
uncoated
pins.
4.
What is the Z clearance on the robot deck
from the highest impediment (top of the source plate/recipient
plate/wash reservoirs, etc.) to the top of the robot mounting
plate? If it is greater than 77 mm you can use our our 30 mm
exposed length FP(#T) or the 17 mm exposed length FP(#) series
pins. If it is less than 77 mm, you can only use the shorter
FP(#)C series pins (12 mm exposed pin length pins and the E-clip series
pins (23 mm exposed pin length.. If it is less than 60 mm, we
recommend you contact us. There are a few tricks we can do to
shave off 5 to 10 mm in height.
Links
to the specifications of all the robot
mounting plates/pin tool/pin combinations can be found on the
height
("Z")
restriction page.
5.
Is the source plate a deep well
plate? If so you may need to use a 30 mm long exposed length
pin ("T" pin)
to
reach down to the lower levels of the well.
6.
Should you choose the solid pin
versus the slot pin? Although the there appears to be a slight
advantage for slot pin, CV's between the two pins
are very good.
Both are easy to clean between specimens. Biggest factor is
cost. If you don't need to deal with varying liquid heights
and
you can obtain the volume necessary with a solid pin, choose the
economical solid pin.
7.
If you are still uncertain about which pin to select for your
application, you can perform a simple
"Proof
of
Principle" test with several
different pins using our inexpensive
VP
450FP3 Replicator
Strip coupled via a
VP
452MP to one of our
robot mounting
plates. An even
simpler solution is to use a work station
with a 1, 4 or 8 probe dispense head with the new
V&P Mono
Pin Tool to test
the various pins.
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Originally
we sold our pin tools with unique part
numbers. As we have added more pins to our selection and more
robot systems to mount the pin tools to, the number of unique part
numbers grew exponentially to an un-manageable size. Our
solution
has been to specify a pin tool by the 3 component parts that compromise
the Pin Tool. The Pins, the Floating Fixture and the Robot
Mounting Plate. If you have previously ordered a pin tool by
unique part number, we will be able to provide the same pin tool to you.
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A
Protedyne Corporation pin tool consists of two
separate parts.
1.
The
pins,
(#FP___)
2. The
Floating
Frame Fixture (384 or 1536)
(#AFIX___FP_)
For
example the part numbers for pin tool in the photo to
the right would be FP1, AFIX1536FP1
To
order, just tell us those 2 part numbers and we will
assemble your Pin Tool.
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1.
Choose
the pin to
match your application, pin length, delivery volume requirements and/or
the non-specific binding nature of the sample.
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Solid pins are the most economical and control volume
transferred by pin diameter.
Solid pins for Protedyne
Corporation are available in 2 exposed pin
lengths; 12 and17mm. The 12 mm ("C") pins are used with a 1536
and the 17 mm ("N") pins are used when
transferring from a 384 or 1536.
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Slot
pins have several advantages over solid pins:
-
Slot
pins carry a larger volume than solid
pins in liquid to liquid transfers.
-
Slot
pins are not as sensitive to the depth
of the mother liquid as solid pens.
-
Slot
pins deliver a larger volume than solid
pins in liquid to dry plate transfers.
-
Slot
pins deliver a larger volume than solid
pins in liquid to membrane transfers.
-
Slot
volume is customizable.
-
Slot pins can deliver to dry plates
by pre-wetting the plate.
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Hydrophobic/lipophobic
coated pins are
very useful in handling
solutions with proteins or lipids that may stick to naked stainless
steel. Both solid and slot pins can be coated.
Click here for more
details on the
nature of
our hydrophobic/ lipophobic coating and the coating process.
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The tables below contain
hyperlinks to the various types/lengths/diameters/coatings of the pins
we offer.
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Please visit
this
hyperlink for Pin Tool Data
And Charts on hydrophobic pins.
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2.
Choose the Floating Fixture to match your
application. Standard are 384 and 1,536 formats.
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These floating fixtures
consist of an upper float plate that
will all accommodate a 1.58 mm diameter tube, a lower float plate that
will accommodate either a .457 mm or .787 mm
diameter pin and 6 standoffs that keep the two plates
separate.
The length of the standoffs is determined by the length of the pin
used. Your application will determine if you select the 384
or 1,536 formats and your previously made pin selection will determine
the diameter of holes in the lower float plate.
Click here
for a
complete selection of these floating fixtures..
If you have
more specific needs call us for help at
(800) 455-0644.
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Call Protedyne Corporation
for pricing information
and tell them the part numbers you need for
the pin tool - Pin # (FP___)
Floating Frame Fixture # (AFIX__FP__ )
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Replacement
Pin Link for the Tube
style Molcular Devices Pin Tools.
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The
Double Float Plate Molecular Devices Tube Style Pin Tools have several
advantages
over
the E-Clip
Style Pin Tools:
-
The
Tube Style Pin Tools are smaller in
diameter and deliver very small volumes (down to 2 nl and as much as
500 nl depending on diameter and slot size).
-
Smaller
pin diameters also result in more
precise delivery volumes (CV's ~4%).
-
Because
the pin is registered between 2 float plates over a greater height, the
pin angle will be truer (closer to 90°)
when they touch
the surface.
-
The
smaller hole on the lower float plate allows us to make very tight
tolerances on the fit between the pin and the hole.
This
results in very precise placement of the pins on the surface so that
very high density arrays can be made without overlapping the spots.
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Custom
Floating Slot Pin Replicators can also be made using the 0.457 mm and
0.787 mm diameter pins and placing them into the Floating Pin
Replicators
described above. The slot volumes between these two pins can
cover a range
of volumes from 5 nl to 1000 nl
by changing the width and depth of the slot. Inquire by phone
or
e-mail
for more information.
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The
most important aspect of using pin tools
is to start with clean pins and then keep them
clean so they will consistently deliver the same volume. We
recommend our Pin
Cleaning Solution (VP
110) to clean the pin of protein
and other residue buildup before
you start an assay. To quickly and efficiently remove the
previous transfer from the pin we have several different
wash
reservoirs, flowing
reservoirs
and fountains to
clean 384 and 1536 floating pin
tools.
These wash stations also are adapted so the
blotting
station
is part of the
washing station thus reducing the cleaning cycle time and the number of
robot
stations required for the cleaning process. We also have
other independent
blotting stations (VP 540D).
Integral to the cleaning process of all pins is the blotting
step between baths. Lint particles in the slots or on pin
tips,
from standard paper towels, will interfere with the loading and
unloading of
liquid. We have solved this problem. The
VP
540DB
blotting system is a tray with the standard microplate foot print that
contains a super absorbent polypropylene pad covered by a
Lint Free Blotting Paper. We strongly recommend that
you use this or our other
lint free blotting systems.
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We highly recommend that you order the Pin Tool Accessory kit
at the time you order the pin tool. This "essentials" accessory kit
contains the Docking Station for the Pin Tool on the robot deck giving you the
capability for "Hot Swapping" the pin tool for other accessories, The VP 540DB Lint Free Blotting
System is used to clean the pins efficiently between samples, two VP 540 Wash
Reservoirs to wash the pins between source plates, the VP 110 Pin Cleaning
Solution is used the clean the pins daily, the VP 425 Pin Cleaning Brush is used
if a build up occurs.
Although we have several
other wash and blot accessories
available these are the most cost effective for the majority of applications.
If you elect to use the fountain and flowing reservoirs we still strongly
recommend the other items in this kit.
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Pin Tool
Accessory
Kit - VP
OK22
|
| Pin
Tool Docking Station for "hot swapping" (1) |
VP 550 |
| Lint
Free Blotting System with pad (1) |
VP
540DB |
| Pin Wash
Reservoirs (2) |
VP
540 |
| Pin
Cleaning Solution (1) |
VP 110 |
| Pin Cleaning
Brush (1) |
VP
425 |
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