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The various types of pins we
use in our Pin Tools are:
1)
Solid
2) Slot
3) Grooved
and 4)
Bolt
type.
They are usually pressed or screwed into an aluminum base
however some
float
freely in aluminum or delrin frames and others are clamped into
scrabble
block fixtures. Our pins can also be provided with a
hydrophobic/lipophobic
coating to reduce non-specific binding of proteins and lipoproteins. Each pin type uses a slightly different method to
pick up and transfer liquid and each has its own advantages. This
page and the tables were designed to help you understand the
capabilities of the various pins and then pick the best one for your
application. We make a great variety of pin tools and this page
will help you focus on the characteristics that are important in your
application.
When these
pin tools are
used with the appropriate robot they are very effective in HTS applications,
screening for compound activity, making serial dilutions,
replicating libraries, making arrays on membranes or slides or
reformatting libraries. See
our guide for selecting the appropriate pin for your robot
application.
We have
made adaptor mounting plates that allow us to mount our pin tools on
34 different commercial robotic
liquid handling robots and we have also made numerous custom mounts
for customer built robotic systems. In many other cases our
customers have purchased the pin tool from us and had their own machine
shop build an adaptor mount. Whatever way you
select, it is simple solution.
If you don't have a robot you can purchase the new
V&P Pin Tool Robot and attach any of our pin tools to it. This
gives you the advantages of robotic reproducibility at a fraction of the
cost of large robotic system.
Our manual pin tools and replicators
when used with the appropriate registration or indexing tool (Glass
Slide Indexer,
MULTI-PRINT™,
LIBRARY COPIER™,
COLONY COPIER™
or Vicki System)
are also very effective in making arrays on slides or membranes, replicating
or reformatting libraries, screening compounds or making serial
dilutions. |
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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. We also tell you
how
to use sonication and
brushes
to clean the pins. To quickly and efficiently remove the
previous transfer from the pin we have several different
wash reservoirs,
flowing reservoirs
and fountains to clean 96, 384 and 1536 floating pin tools.
These robot 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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By
simply dipping a solid pin into a liquid and removing it, you
get a very uniform droplet of liquid that hangs on the tip of
the pin. This droplet is very uniform in volume.
Its
size depends on several factors: 1) Pin diameter 2) Shape
of the tip {flat, chamfered or pointed} 3) Surface tension on the pin 4) Surface
tension of the liquid and 5) The speed at which the pin is
removed from the liquid. 6) The depth to which the pin is
submerged. If all these factors remain the
same, the size of the drops will be very precise. Thus for
applications where you are just transferring the hanging drop to
a membrane or agar surface, you can get accurate and
precise delivery of liquid. If the delivery is to another
liquid, then the sides of the pins which also collect liquid
droplets will be added to the delivery. In addition, the
total volume transferred in liquid to liquid transfers is also
dependant upon how far the pins are submerged in both the mother
wells and the daughter wells. As long as the pin transfer
is done in a consistent manner you will achieve
very
good CV's.
If you are transferring the hanging drop to a dry well then the
speed of the pin striking the surface is a factor in how much
gets transferred as well as the surface tension of the dry well.
As a general rule of thumb about
35% of the hanging drop is transferred to the dry well.
With robot controlled delivery speed good CV's are obtained. |
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The advantage of the solid pins is that the
hanging drop provides a very accurate and precise method of
transferring as little as 2 nanoliter volumes of liquid to membranes
or dry microplate wells.
The following table organizes our Fixed Solid Pin
Replicators according to:
-
Volume of the hanging drop (this is an
approximate value as their are a number of characteristics
such as liquid surface tension that will affect the volume).
-
Pin diameter
-
Pin number
-
Pin type (Fixed, Long or Scrabble)
-
Catalog number
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Hanging
Drop |
Pin
Diameter |
Pin # &
Type
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Catalog # |
Pin Part # |
| ~20 nl |
0.457 mm |
384 Fixed |
VP
384VR1 |
P1 |
| ~20 nl |
0.457 mm |
1,536 Fixed |
VP 838 |
P1 |
| ~100 nl |
1.19
mm |
96
Fixed |
VP
409 |
P2 |
| ~100 nl |
1.19
mm |
384
Fixed |
VP
386 |
P2 |
| ~200 nl |
1.58
mm |
48
Fixed |
VP
480 |
P |
| ~200 nl |
1.58
mm |
96/2
Fixed |
VP
408H |
P |
| ~200 nl |
1.58
mm |
96
Fixed |
VP
408 |
P |
| ~200 nl |
1.58
mm |
96 Long |
VP
408A |
PA |
| ~200 nl |
1.58
mm |
96
Scrabble |
VP
401 |
P |
| ~200 nl |
1.58
mm |
384
Fixed |
VP
384 |
P |
| ~1.5 ul |
2.36
mm |
96
Fixed |
VP
407 |
P12 |
| ~3 ul |
3.18
mm |
96
Fixed |
VP
407A |
P13 |
| ~3 ul |
3.18
mm |
48 Fixed |
VP
407AH |
P13 |
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4.75 mm |
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P11 |
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Fixed pins have been
press fit into an aluminum or delrin base.
Pin
Tool Data And Charts
For
customer pin tool data please visit this hyperlink
Detailed
Assay Method:
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Floating pins have several
advantages. First, the pins are all assured of touching an uneven
surface such as agar, a membrane or microplate with a warped bottom
which is common with polypropylene microplates. Secondly,
individual pins with different delivery volumes can be selected and
arranged by the user in the Replicator to facilitate making serial
dilutions in the same Replicator. Like wise damaged pins are
easily replaced. The floating design can be used
manually or also
by robots where the Z control may vary.
We have two styles of
Floating Pin Replicators: The
E-Clip
style which is the least expensive and transports the largest volume
of liquid. The
Tube style which
comes in 5 different pin diameters and transports the smallest volume of liquid with very
precise pitch location and absolute perpendicular vertical alignment.
The Tube style also comes in three different lengths. The
standard tube
style length is 50.8 mm long with a 17 mm long exposed pin length.
This is the maximum distance that the pin will stick into the microplate
well. On some robot workstations there are height limitations and
for those robot systems we made a
shorter pin (FPCB series)
that is 33 mm long with a 12 mm
exposed pin length. For applications using deep well
microplates you need a pin with a longer exposed length to reach farther
down into the wells. The
FPT series pins have pins with 30 mm exposed pin lengths to adapt to
these applications.
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Long pins are for use
on deep well plates, they are 45 to 57 mm in length and may be
slotted
or solid.
Scrabble
Block is a type of fixed pin base that allows the customer to change
pins between assays or to mount delicate pins that cannot be
pressed. With the Scrabble base it is possible to
arrange slot pins of different volumes in what ever location
that fits the experiment. |
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The FP series Solid
and Slot Pins are also available with
a hydrophobic/lipophobic coating. This coating prevents or
reduces the amount of non-specific binding of proteins and lipids to the
pins. It will also reduce "creep" of liquid up the pin. The hydrophobic/lipophobic
versions are designated with an H in the catalog number. They can be
found on the
Pin page. |
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By simply
cutting slots into the tips of solid pins, the slots will fill
precisely, by capillary action, and then deliver that
amount by capillary wicking onto a membrane or by immersion into
another liquid. Usually the volume in the slot is
significantly more than the drops on the side of the pin so the
precision of liquid transfer is not effected greatly in liquid
to liquid transfers and not at all in liquid to membrane
transfers. One of the phenomena of slotting pins is the
great reduction in the hanging drop at the tip of the pin
because of the slot. This results in the volume of liquid
to be transferred, being close to the volume of the slot.
Note that slot pins will not
transfer the volume of the slot to a dry well or glass slide but
can be made to deliver a larger volume than a hanging drop by
causing the pin to strike the surface at a high rate of speed.
This is best achieved when controlled by a robotic system.
Because it is relatively easy for us to cut any size of slot we
choose, it is simple to make custom Slot Pin Replicators.
We have several that we make and stock and they are listed
below. If you don't see what you need, call us and we will
make a special one for you.
The advantage
of the slot pins is that the slot provides a
very
accurate and precise method of transferring 10 nl
and greater volumes of liquid to membranes or to liquid in
daughter wells. They are also useful in spotting nanoliter
volumes onto glass slides.
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FP4 slot pins are our newest slot pins.
Although they transfer relatively large volumes (1,000 and 2,000 ul)
that could be easily handled by the 1.58 mm diameter pins, they are
the largest diameter slot pins that can still fit inside the wells in
a 1,536 well microplate. Thus companies that have their compound
libraries in 1,536 well microplates find them very useful in High
Throughput Screening assays done in the 1536 format. |
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The larger slot volume FP6 series floating pins cover the slot
volumes from 0.5 to 5ul. |
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These replicators were made in response to scientists
that want to deliver larger inoculums (usually yeast) to an agar
Omni Tray or a
membrane. To make slots that will yield the larger volumes we have
to use pins of a larger diameter. The end view of the slot is
illustrated in photo to the right. The 25 ul P11AS25 pins are 4.75
mm in diameter and the 10 ul P13S10 pins are 3.17 mm in diameter. |
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We can also
make custom slot pins
that will deliver as little as 6 nl to meet your specific
needs.
Also see our 8, 12, 16 and 24
Replicator Strips with slot pins
and 8, 12, 16 and 24
Floating Replicator Strips with slot pins.
These are good for loading gels and making microarrays on glass slides.
The 22 mm long pins are better for loading MADGE
Gels or inoculating onto an agar surface. Both of the pins are
used to make quantitative liquid to liquid transfers or liquid to membrane
blots.
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Pin
Tool Data And Charts
For
customer pin tool data please visit this hyperlink
Detailed
Assay Method:
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Grooves
at the end of a solid pin will fill precisely by capillary
action just as the slots do. Grooved pins are only used for
liquid to liquid transfers.
The
advantage of grooved pins is that they provide a very accurate
and precise method of transferring from 3ul
to 90ul. |
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| ~Groove
Volume |
Pin
Diameter |
Pin # &
Type
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Catalog #
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| 3 ul |
2.77
mm |
96 Fixed, 22 mm long |
VP
403D1 |
| 3 ul |
2.77
mm |
96 Fixed, 22 mm long |
VP
403D1a |
| 3 ul |
1.58
mm |
96 Fixed, 57 mm long |
VP
408B |
| 5 ul |
2.77
mm |
96 Fixed, 13 mm long |
VP
403B |
| 5 ul |
2.77
mm |
96 Fixed, 22 mm long |
VP
403D2 |
| 6 ul |
2.77
mm |
96 Fixed, 47 mm long |
VP
403F |
| 7 ul |
3.18
mm |
96 Fixed, 22 mm long |
VP
403A |
| 20 ul |
3.18
mm |
96 Fixed, 22 mm long |
VP
403E |
| 50 ul |
6.35
mm |
24 Fixed, 57 mm long |
VP
224A |
| 90 ul |
6.35
mm |
24 Fixed, 57 mm long |
VP
224B |
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The threads of the bolt pick up large volumes of liquid for liquid
to liquid transfers or for spotting yeast onto agar plates. The volume of
liquid transfer depends upon the pitch and depth of the threads and the
depth to which the threads are submerged in the mother and daughter wells.
The Bolt Type Replicators are used where precision of transfer is not
important.
Their advantage is that they are
inexpensive and transfer larger volumes. |
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| ~Thread
Volume |
Bolt
Specifications |
Bolt
# & Length |
Catalog
# |
| 10 ul |
8-32 |
96,
18 mm long |
VP
404 |
| 10 ul |
6-32 |
48,
17 mm long |
VP
404A |
| 20 ul |
10-24 |
96,
17 mm long |
VP
405 |
| 50 ul |
8-32 |
96,
53 mm long |
VP
404B |
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We also provide many of the same pins as
Replicator
Strips (a single row or column of 8, 12, 16 or 24 pins arranged in
96 or 384 well format - 9 mm or 4.5 mm centers). |
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We also provide some of the same pins in
2 and 4 row strips for spotting DNA onto glass slides in high density
arrays. Glass Slide Replicators for 96 well microplates and 384 well
microplates are found on the
Glass
Slide Arrayer page. |
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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 robot applications the CV are less than 5%.
Probably the most useful factor is that by
robotically changing the speed of withdrawal from the source plate you
can precisely determine the volume of liquid on the pin over a large
dynamic range. This range is illustrated in the table below.
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FP9 |
0.229 mm |
Solid |
5
- 15 nl |
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FP8 |
0.350 mm |
Solid |
12 – 28 nl |
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FP1 |
0.457 mm |
Solid |
19 – 68 nl |
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FP1S6 |
0.457 mm |
6 nl Slot |
26 – 74 nl |
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FP1S10 |
0.457 mm |
10 nl slot |
31 – 79 nl |
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FP1S20 |
0.457 mm |
20 nl slot |
43 – 89 nl |
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FP1S30 |
0.457 mm |
30 nl slot |
53 – 99 nl |
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FP1S40 |
0.457 mm |
40 nl slot |
64 – 109 nl |
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FP1S50 |
0.457 mm |
50 nl slot |
74 – 117 nl |
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FP3 |
0.787 mm |
Solid |
84 – 240 nl |
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FP4 |
0.910 mm |
Solid |
112 – 207 nl |
| FP |
1.580 mm |
Solid Pointed |
157 – 595 nl |
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FP3S100 |
0.787 mm |
100 nl slot |
178 – 326 nl |
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FP2S200 |
0.787 mm |
200 nl slot |
278 – 424 nl |
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FP6 |
1.580 mm |
Solid Flat |
363 – 884 nl |
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FP3S500 |
0.787 mm |
500 nl slot |
527 – 656 nl |
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FPS.5 |
1.580 mm |
0.5 ul slot |
443 – 906 nl |
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FP6S.5 |
1.580 mm |
0.5 ul slot |
726 – 1,253 nl |
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FPS |
1.580 mm |
1 ul slot |
938 – 1,424 nl |
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FP6S |
1.580 mm |
1 ul slot |
1,239 – 1,786 nl |
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FPS2 |
1.580 mm |
2 ul slot |
1,817 – 2,248 nl |
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FP6S2 |
1.580 mm |
2 ul slot |
2,102 – 2,576 nl |
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FPS5 |
1.580 mm |
5 ul slot |
4,475 – 4,828 nl |
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FP6S5 |
1.580 mm |
5 ul slot |
4,475 – 4,828 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
The links below are to delivery volume range tables for
uncoated and hydrophobic coated delivering either DMSO or Aqueous solutions
in liquid to liquid transfers.
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 any of pins. If it is less than 77 mm, you can
only use the shorter FP(#)C series pins (12 mm exposed pin length), the FP(#N)
series pins (17 mm exposed pin length) and the
E-clip series pins (23 mm exposed pin length pins. If the Z clearance 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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Many of our customers want to make liquid to dry
plate transfers to make mini-microarrays in a single well for
multiplexing assays. By their very nature these transfers only leave
a small volume remaining on the plate. The amount remaining
depends on the surface tension of the plate, the transfer liquid and
the pin (diameter and shape). We have found that reasonable results can be obtained
if you use an aqueous spotting buffer and a polypropylene surface
(CV's less than 9%). DMSO spotting buffers
leave much less liquid on the dry plate and when used with
polystyrene plates the
CV's are significantly
higher.
The plate to the right was spotted using a 50 nl
slot pin (FP1CS50) transferring oligo DNA in an aqueous spotting
buffer to a specially treated polypropylene plate. Our customer in this case wanted to place 42 individual
spots in the bottom
of each well
of a 96 well microplate. Spot diameter =
0.55 mm. Volume transferred to the spot = 5.85 nl The photo on the right is the result of a
four fold serial
dilution hybridization assay of the Oligo spots with a labeled DNA
that can be detected optically and quantitated.
Follow this link for more information on
liquid to dry transfers and
Multiplexing. |
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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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1.
Select the appropriate pin.
For the manual and Beckman Bio Mek 2000 robots we recommend the longer FP(#)
series pins.
2. Select the
Floating Fixture with the
appropriate pattern (96, 384 or 1,536) and diameter of holes. |
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3. Select the robot system you wish to use the Pin Tool
with. We have made
Mounting Plates
to over 34 Robot Workstations.
Determine the weight of a Robot pin tool at
this link |
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Our Robotic floating Pin Tools can easily be converted to
a manual pin tool with this simple
BGPK kit
which contains a
Guide Pin Frame and handle and sturdy mounting plate
(BMPM) The
guide pins are used with our manual microplate (VP 381N) and membrane registration
accessories (VP 382 and VP 382B) to
keep the pins in the center of the wells and to
make high
density arrays on membranes or agar surfaces.
Conversion of robot pin tools to manual Multi-Blot Replicators is very
easy using our BGPK for Standard length pins and the BGPKL for
longer pins. Simply remove the robot mounting feature from the float
plates disassemble the pin tool and reassemble it with the Guide Pin Frame
as part of the tool. Attach the Manual Mounting Plate and handle and
you are ready to go. If this sounds too daunting for you - send
it back to us and we will do it for you for a nominal fee. |
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The main reason for this conversion is to attach
guide pins to the replicator so it can be aligned to Microplate registration
accessories such as the VP 381 series Library copiers for replicating and
reformatting genomic libraries, VP 380 series colony copiers to make high
density arrays on agar surfaces or VP 382 membrane printers to make
high density arrays on nylon or other membranes.
The
BGPKL
kit is used when longer replicator pins are used like the
FPAM pins
which are 63 mm long. A pin tool with FPAM pins is
illustrated in the photo to the right. In order to accommodate
these longer pins we need to use a longer and more sturdy guide pin found on
the BGPL frame. These larger guide pins are 3.17 mm in
diameter.
Each of the kits on the right (BGPKL and
BGPK) is made up
of a handle, a manual mounting plate and a guide pin frame. The guide
pin frame is the only difference between the two kits
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