Agilent Technologies PlateLoc User manual


PlateLoc Thermal
Microplate Sealer
Quick Reference Guide

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G5402-90003
June 2009 rebranded edition
May 2008 quick reference guide edition
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,

1
PlateLoc Thermal Microplate Sealer
Reference for Optimal Thermal Microplate Sealing

PlateLoc Thermal Microplate Sealer
Reference for Optimal Thermal Microplate Sealing
This reference contains the following topics:
•“Optimizing the sealing quality” on page 2
•“Optimizing seal parameters” on page 5
•“Requesting application support” on page 8
•“Frequently asked questions” on page 9

PlateLoc Thermal Microplate Sealer
About this guide
2Reference for Optimal Thermal Microplate Sealing
About this guide
This guide provides a quick- reference for using the PlateLoc Thermal
Microplate Sealer (PlateLoc Sealer). Use the following guidelines to identify
thermal- sealing- friendly labware and optimize microplate sealing using the
PlateLoc Sealer. You should use this guide in conjunction with the following:
•PlateLoc Thermal Microplate Sealer User Guide
•PlateLoc Seal Selection Guide
You can download the latest version of any PDF file or search the online
knowledge base from the Automation Solutions Knowledge Base page.
For more information about automation products, go to the Agilent
Technologies website at www.agilent.com/lifesciences/automation.
Optimizing the sealing quality
Introduction
The PlateLoc Sealer applies a sealing material on top of microplates to seal
individual wells. Sealing the wells protects the contents from evaporation,
condensation, oxidation, and cross- contamination during transport or storage.
The PlateLoc Sealer can be used as a standalone device or in a lab automation
system. The device accepts microplates made from a variety of materials.
The ideal sealing conditions can depend on a number of factors, especially the
labware. Using thermal seal- friendly microplates is key to protecting samples
and critical to successful sample sealing and storage applications.
Microplate sealing tips
The seal quality depends on many factors, including microplate design,
material, lot- to- lot consistency and features, the sealing parameters, whether
the microplates require an insert for support, and the type of seal used.
Microplate design
Automation- friendly and thermal- seal- friendly labware ensures successful
sealing.
• Use only microplates that comply with standards established by the Society
of Biomolecular Sciences (SBS).
• Raised chimneys around each well produce the best results.
• Avoid microplates that have a raised rim close to the perimeter of the
wells. The rim can prevent uniform contact between the hot plate and the
wells.
Figure 1 illustrates these features.

3
PlateLoc Thermal Microplate Sealer
Optimizing the sealing quality
Reference for Optimal Thermal Microplate Sealing
Figure 1 Microplate physical features and nomenclature
Figure 2 Left: Pneumatics drive the hot plate down to the microplate surface. The hot plate can
sense and automatically adjust for variations in microplate height. Seals are applied using
pressure and heat. Right: If the microplate has a raised rim that is higher than the chimney height
around its perimeter, the hot plate must fit within the rimmed area so that the hot plate can make
uniform contact with the seal and all sample wells simultaneously.
Microplate material
For the best sealing results:
• Use polypropylene microplates because they are the easiest to seal
effectively and reliably.
• Avoid acrylic microplates because they are not compatible with thermal
microplate sealing.
• If the microplates are flexible and tend to bend and move during the
sealing process, use adapter trays to stabilize the microplate position on
the plate stage. For example, place a flexible 96- well polypropylene
microplate on top of a 96- well polystyrene microplate to provide positional
support. Agilent Technologies also offers an adapter tray for PCR
microplate support.
Raised rim around
perimeter (if any) Sample well
Skirt
Plate
surface
Chimney: Top of well rises
beyond microplate surface
Insert
HOT PLATE
HOT PLATE
Microplate with
raised rim
Sealing film
Hot plate

PlateLoc Thermal Microplate Sealer
Optimizing the sealing quality
4Reference for Optimal Thermal Microplate Sealing
Figure 3 Left: Microplate and PCR Plate Support. Right: Microplate in PCR Plate Support.
Microplate make and model
• Be aware that similar microplates made from the same material but from
different manufacturers may perform differently.
• Evaluate every microplate make and model to ensure consistent results.
Microplate features
Be aware of design features on the top surface of the microplate that can
significantly affect the quality of the seal.
• The microplate lacks raised chimneys and it has molded branding or
legends, such as the manufacturer’s name or logo, or the sample locating
legends (for example A, B, C, . . . and 1, 2, 3 . . .) close to the tops of the
wells (close enough so that seal touches or covers the imprint).
• Tooling marks or manufacturing aids, such as circular depressions
(sometimes with branching) caused by ejection pins or part of the molding
process. Gaps or gutters may not only prevent binding of the seal, but may
actually facilitate cross- contamination of liquid samples.
Sealing parameters
The following parameters can be adjusted to optimize the sealing results:
•Temperature
•Time
Perform optimization tests to determine the best values to use. For details, see
“Optimizing the sealing parameters (temperature versus time)” on page 10.
Insert thickness
Choose the right insert to support the microplate and ensure uniform sealing.
A set of four inserts are presently shipping with every PlateLoc Sealer, and a
special insert is available for Labcyte microplates.

5
PlateLoc Thermal Microplate Sealer
Optimizing seal parameters
Reference for Optimal Thermal Microplate Sealing
Figure 4 Microplate without and with insert
Figure 5 Standard inserts. The number on the inserts indicates the thickness of the metal pad
(for example, 180 means 0.180 in thick). Note: The 90 insert has a 0.90-in metal pad with foam
padding on the microplate-facing side and is used with microplates that require flexible support.
Seal material
Different seal materials have different properties and can produce different
results. Perform optimization tests to determine the best seal to use.
Optimizing seal parameters
About this section
This section explains how to inspect the seal quality and, if necessary, how to
adjust the sealing parameters to improve the seal. Before proceeding, download
a copy of the PlateLoc Seal Selection Guide from the Agilent Technologies
website at http://www.chem.agilent.com/en-us/products/instruments/automation/
microplatehandling/plateloc/pages/default.aspx.
Apply a seal using the time and temperature starting points suggested in the
PlateLoc Sealer Selection Guide, for your chosen seal and microplate material.
Examine the well impressions made on the seal. The pattern and quality of
these impressions is a good indicator of seal quality.
Evaluating the seal
After the seal cycle is finished, wait for the microplate to cool down enough so
that it is safe to handle (10 to 30 seconds). Do not handle the microplate
immediately after the seal cycle is finished. The sealed microplate might be
hot.
Note: Pierceable seals might permanently adhere to the microplate. Do not wait
too long or the seals will be difficult to remove.
Remove the microplate from the PlateLoc Sealer. Carefully peel off the seal
material by lifting one of the corners of the seal as shown in Figure 6.
Bend direction
Microplate insert
Wells

PlateLoc Thermal Microplate Sealer
Optimizing seal parameters
6Reference for Optimal Thermal Microplate Sealing
Figure 6 Removing the seal.
Inspect the underside of the removed seal material. If there are unbroken
impressions of each well on the underside of the seal material, the microplate
was properly sealed.
Figure 7 Example of a good seal.
Use a magnifying glass or inspection microscope, if necessary, to see the area
surrounding the sample well rim.
Look closely at a single sample well. A good seal might look like the
illustrations on the left side of Figure 8. The well impression should match the
thickness of the chimney wall, as shown in the left side of Figure 9. A poor
seal might look like the illustrations on the center or right side of Figure 8,
indicating there may not have been enough heat applied to form a complete
impression of the sample well rim.
Figure 8 Evaluating the seals of sample wells. L: Good impression, with defined edge of
chimney/rim. C and R: Poor impression, with missing or very faint segments.
If there are faint or broken impressions in the seal material, as shown in
Figure 8, increase either the sealing temperature or time (duration).
00201 PlateLoc sealed plate
00201 PlateLoc sealed plate

7
PlateLoc Thermal Microplate Sealer
Optimizing seal parameters
Reference for Optimal Thermal Microplate Sealing
If the well impressions on the seal are significantly thicker than the chimney
wall (as shown in Figure 9), decrease the sealing temperature or duration.
Thick impressions may indicate excessive melting of the seal, causing the seal
to be difficult to remove and the resealing capacity reduced.
Figure 9 Evaluating rim width of seal. L: Good impression, with thin, defined edge of chimney/
rim. R: Poor impression, with increased width.
Issues related to inserts
The hot plate is a heated metal surface that presses the seal onto the
microplate. For optimal sealing, the microplate needs to be firmly supported so
that it does not bend during the sealing process, often requiring use of an
insert to keep it flat. Figure 10 illustrates the effect of insert height on
microplate sealing. Left: if the insert is too thin, the center of the microplate
will bend downward during sealing, leading to good sealing around the edges,
but poor sealing in the middle of the microplate. Center: if the insert is the
correct thickness, the entire microplate surface will be sealed properly. Right:
if the insert is too thick, the edges of the microplate will bend downward
during sealing, resulting in good sealing in the middle of the microplate, but
poor sealing around the edges.
Note: Thicker inserts are preferred over inserts that are too thin.
Figure 10 The effect of using inserts on microplate sealing. L: Insert too thin; C: Insert correct
height; R: Insert too thick.
Approximately same
thickness as chimney
wall
Thicker than chimney wall

PlateLoc Thermal Microplate Sealer
Requesting application support
8Reference for Optimal Thermal Microplate Sealing
Requesting application support
To help expedite your support request, you can complete and submit the
Request for PlateLoc Sealer Application Support.
For other questions, you can contact Automation Solutions Technical Support
at one of the following:
Europe
Phone: [phone]
email: [e-mail]
US and rest of world
Phone: [phone] (US only) or +1.408.345.8011
email: [email protected]