Thermo Scientific mDrop User manual

Cat. No. N12691 Rev 4.0 2020
Thermo Scientific
μDrop and μDrop Duo
Plates
User Manual

Copyright
Copyright © 2020 Thermo Fisher Scientific Inc. All rights reserved. Reproduction of the accompanying user
documentation in whole or in part is prohibited
Manufacturer
Life Technologies Holdings Pte. Ltd.
(a part of Thermo Fisher Scientific Inc.)
33, Marsiling Industrial Estate Road 3, #7-06,
Singapore 739256
Trademarks
“μDrop” is a trademark of Thermo Fisher Scientific. “Multiskan”, “SkanIt” and “Varioskan” are registered
trademarks of Thermo Fisher Scientific.
All other trademarks and registered trademarks are the property of their respective holders.
Disclaimer
Thermo Fisher Scientific reserves the right to change its products and services at any time to incorporate
technological developments. This manual is subject to change without prior notice as part of continuous
product development. Although this manual has been prepared with every precaution to ensure accuracy,
Thermo Fisher Scientific assumes no liability for any errors or omissions, nor for any damages resulting from
the application or use of this information. This manual supersedes all previous editions.
No liability for consequential damages
Thermo Fisher Scientific shall not be liable for any damages whatsoever arising out of the use or inability to use
this product.

Thermo Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual i
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Preface
About This Guide
This manual gives instructions on performing absorbance measurements using the Thermo Scientific™
μDrop™ and μDrop™ Duo Plates with the following instruments:
• Thermo Scientific™ Multiskan™ SkyHigh
• Thermo Scientific™ Multiskan™ Sky
• Thermo Scientific™ Multiskan™ GO
• Thermo Scientific™ Varioskan™ LUX
Safety and Special Notices
Make sure you follow the precautionary statements presented in this guide. The safety and other
special notices appear in boxes.
The following symbols and markings appear in this user manual:.
WARNING Biohazard risk.
WARNING Risk of injury to the user or users.
CAUTION Risk of damage to the instrument, other equipment or loss of performance or function
in a specific application.
Note Highlights important information, a tip, or an item of interest that is useful in the optimum
operation of the system

About This Guide
ii Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Scientific
Contacting Us
For the latest information on products and services, visit our website at:
www.thermofisher.com/platereaders

Thermo Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual iii
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Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
About This Guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Safety and Special Notices. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Contacting Us. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii
Chapter 1 Introduction to the μDrop and μDrop Duo Plates . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
General description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Chapter 2 Low-volume measurements with SkanIt Software . . . . . . . . . . . . . . . . . . . . . . . . . 5
Creating a new measurement session . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Ready-made sessions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
User-made sessions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Placing the samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Cleaning verification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Chapter 3 Cuvette measurements with SkanIt Software. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Placing the cuvette. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Creating a new measurement session . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
User-made sessions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Chapter 4 Setting up a measurement from the instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Chapter 5 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Regular and preventive maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
μDrop and μDrop Duo Plates care . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Cleaning the μDrop and μDrop Duo Plates. . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Disposal of materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Decontamination procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Disposal of the μDrop and μDrop Duo Plates . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Chapter 6 Technical specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
General specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Contents

Contents
iv Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Scientific

Thermo Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual 1
1
Introduction to the μDrop and μDrop Duo Plates
Introduction
Thermo Scientific™ μDrop™ and μDrop™ Duo Plates are tools designed for quick and easy absorbance
measurement of low sample volumes (2–10 μl) and cuvettes with the following instruments:
• Thermo Scientific™ Multiskan™ SkyHigh
• Thermo Scientific™ Multiskan™ Sky
• Thermo Scientific™ Multiskan™ GO
• Thermo Scientific™ Varioskan™ LUX
The μDrop and μDrop Duo Plates are typically used for photometric DNA, RNA and protein
quantitation and purity analysis.
The μDrop Plate consists of two separate measurement locations, one for measuring low-sample
volumes and the other for cuvettes. The μDrop Duo Plate consist of three separate measurement
locations, two for measuring low-sample volumes and one for cuvettes.
The low-volume area of the μDrop and μDrop Duo Plates is compatible with an 8-channel pipette to
enable easy pipetting of the samples to the μDrop and μDrop Duo Plates. It can be used for endpoint
and spectral measurements.
The cuvette slot is compatible with standard rectangular cuvettes with a stopper. It can be used for
endpoint, spectral and kinetic measurements.
Intended use
The μDrop and μDrop Duo Plates enable quick and easy absorbance measurements with low sample
volumes and standard rectangular cuvettes with a stopper. The main applications are photometric
nucleic acid and protein analysis.
The μDrop and μDrop Duo Plates are intended to be used in research laboratories by professional
personnel.
The μDrop and μDrop Duo Plates are designed to be a part of an analyzing system for the end user,
who is responsible for validating the system to ensure reliable and safe results. We recommend using
Good Laboratory Practice (GLP) during the analysis process.

1Introduction to the μDrop and μDrop Duo Plates
General description
2Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Scientific
General description
The μDrop Plate low-volume measurement area on the left-hand side consists of two quartz slides, the
top clear quartz and the bottom Teflon-coated quartz slide (see Figure 2). The bottom slide contains 16
sample positions, arranged in a 2 x 8 matrix, where samples are to be pipetted. The nominal pathlength
for the sample is fixed to 0.5 mm. The verified pathlength for each μDrop Plate is indicated on the
quality control measurement report delivered with the μDrop Plate.
The μDrop Duo Plate has two low-volume measurement areas on the left-hand side (see Figure 4). The
verified pathlengths for each μDrop Duo Plate are indicated on the quality control measurement report
delivered with the μDrop Duo Plate.
On the right-hand side, the μDrop and μDrop Duo Plates have a slot for cuvette measurement (see
Figure 1 and Figure 3).
Figure 1. A closed μDrop Plate showing the low-volume measurement area (1), adjustment screw (2), and
the cuvette measurement point (3).

1 Introduction to the μDrop and μDrop Duo Plates
General description
Thermo Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual 3
Figure 2. An open μDrop Plate showing the top slide (1), bottom slide (2), sample positions A2 - H3 (2 x 8
matrix) (3), low-volume area (4), and cuvette slot (5).
Figure 3. A closed μDrop Duo Plate showing the low-volume measurement area (1), adjustment screw (2),
and cuvette measurement point (3).

1Introduction to the μDrop and μDrop Duo Plates
General description
4Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Scientific
Figure 4. An open μDrop Duo Plate showing the top slides (1), bottom slides (2), sample positions (two 2 x
8 matrixes) (3), low-volume area (4), and cuvette slot (5).
The measurement sessions for Multiskan SkyHigh, Multiskan Sky, Multiskan GO and Varioskan LUX
can be created using Thermo Scientific™ SkanIt™ Software. It is also possible to measure the μDrop and
μDrop Duo Plates with Multiskan SkyHigh and Multiskan Sky internal software ready-made
protocols.

Thermo Fisher Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual 5
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Low-volume measurements with SkanIt Software
The μDrop and μDrop Duo Plates are used to perform photometric absorbance measurements from
microliter volumes.
Creating a new measurement session
To create new measurement sessions using SkanIt Software:
• Open ready-made sessions.
• Create your own user-defined sessions.
Ready-made sessions
The ready-made sessions for commonly used assays can be downloaded from the SkanIt Cloud Library.
You can access the Cloud Library sessions if you have an Internet connection.
Figure 5. SkanIt Cloud library
For more details about the SkanIt Cloud Library, refer to the Thermo Scientific™ SkanIt™ Software for
Microplate Readers Technical Manual (Cat. No. N16046). It is recommended to visit the SkanIt Cloud
Library to get the latest versions of the sessions, and also any possible new sessions.

2Low-volume measurements with SkanIt Software
Creating a new measurement session
6Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Fisher Scientific
The demo sessions contain different protocols for photometric quantification of nucleic acids and
proteins with representative results. They also contain ready-made calculations for concentrations as
well as purity values. For detailed descriptions and instructions for using a certain session, refer to the
Notes field of the session in question. The sessions can be imported to SkanIt Software and taken into
use either as they are or after modifications. For more details about SkanIt Software, refer to the
Thermo Scientific™ SkanIt™ Software for Microplate Readers Technical Manual (Cat. No. N16046).
Pathlength
To be able to follow the Lambert-Beer’s law for calculating the concentration of nucleic acids and
proteins, the light pathlength of the sample has to be known and it has to be expressed in cm.
The μDrop Plate has a fixed nominal pathlength of 0.05 cm (0.5 mm).
In the case of the μDrop Plate, the pathlength provided in the certificate (in cm) has to be used in the
Lambert-Beer's equation to calculate the concentration of the target molecules.
In the case of the μDrop Duo Plate, two pathlengths are provided (one for each 2x8 matrix), and these
values need to be separately used in the calculations for the unknown samples located in each matrix.
In the provided sessions, example pathlengths (of, for instance, 0.049 or 0.051 cm) are used in the
calculation step of SkanIt Software. For accurate calculations, replace those values with those given on
the quality control measurement report of the individual μDrop or μDrop Duo Plate. After replacing
the pathlength(s), save the modified session.
User-made sessions
To create measurement sessions for the μDrop and μDrop Duo Plates, follow the normal SkanIt
Software procedure. However, there are certain special features to observe:
1. Plate Layout
Use the Thermo Scientific μDrop Plate or Thermo Scientific μDrop Duo Plate plate template in
SkanIt Software. Select the correct template from the Change plate template dropdown menu, see
Figure 6.
Note With Varioskan LUX, use the 96-well adapter for a plate without a lid (ID no. 2).

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Creating a new measurement session
Thermo Fisher Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual 7
Figure 6. Selecting the plate template
All existing sample types are available for the layout (see Figure 7 and Figure 8) and calculations.
Figure 7. Example layout of μDrop Plate containing only blanks

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Creating a new measurement session
8Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Fisher Scientific
Figure 8. Example layout of μDrop Duo Plate containing only blanks
2. Protocol
All photometric measurement types can be used with the μDrop and μDrop Duo Plates. Note that
long kinetic measurements or incubation are not recommended due to evaporation.
The μDrop and μDrop Duo Plates can be used in the whole wavelength range of the instrument in
question.
Figure 9. Protocol example.
Note The μDrop and μDrop Duo Plates support only photometric measurements.
Note The μDrop and μDrop Duo Plates do not support dispensing.
Note Shaking the μDrop and μDrop Duo Plates is not recommended.

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Creating a new measurement session
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3. Results
All available calculation methods can be used similarly to normal microplate measurements.
For the μDrop Plate, the pathlength provided in the certificate (in cm) has to be used in the
Lambert-Beer's equation to calculate the concentration of the target molecules.
For the μDrop Duo Plate, two pathlengths are provided (one for each 2x8 matrix), and these
values need to be separately used in the calculations for the unknown samples located in each
matrix.
For example, for calculations of the dsDNA concentrations (μg/mL), the following formula is
used:
c=(A260-A320)*((50 μg/ml)/PATHLENGTH_CM)
where:
c = dsDNA concentration in μg/mL
A260 = absorbance value at 260 nm after blank subtraction
A320 = absorbance value at 320 nm after blank subtraction
PATHLENGTH_CM = pathlength value of the μDrop Plate in cm
For the μDrop Duo Plate, if different pathlengths are reported for each 2 x 8 matrix, then two
separate calculations of concentrations need to be carried out. To accomplish that, two different
sample groups can be, for instance, created in SkanIt Software (one group for each 2 x 8 matrix).
The calculation is then performed separately for each group (2 x 8 matrix side), with the
corresponding pathlength value.
To add the equation, click Results > Custom Formula (see Figure 10).
Figure 10. Custom Formula
For more details about SkanIt Software, refer to the Thermo Scientific™ SkanIt™ Software for
Microplate Readers Technical Manual (Cat. no. N16046).
Blanks
Each measurement should contain a blank subtraction. It is recommended to use at least one blank
sample in each measurement. There are different ways to perform the blank subtraction:
• Blanks on the same μDrop or μDrop Duo Plate

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Placing the samples
10 Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Fisher Scientific
• Sample specific blanks
•Previouslymadeblanks
Placing the samples
Before starting the measurement, make sure that the μDrop or μDrop Duo Plate is clean after the
previous measurement. For instructions, refer to “Cleaning the μDrop and μDrop Duo Plates” on
page 19.
The samples are pipetted to the sample area using either a single or a multichannel pipette. The sample
volume must be between 2 and 10 μl, 4 μl recommended.
Possible air bubbles present in the liquid can cause severe errors to the results. It is particularly
important to follow good pipetting practices. The use of an 8-channel pipette is recommended. The
reverse pipetting technique is strongly recommended to avoid bubbles.
Because of the very small sample volumes, evaporation is an important issue concerning the reliability
of the result. Therefore, it is important to pipette the samples into the small wells as fast as possible and
perform the measurement as soon as possible after pipetting, especially when using a single pipette.
With smaller sample volumes, a larger evaporation effect can be observed due to changes in
concentration.
1. Open the lid covering the dotted sample area (see Figure 2 on page 3 and Figure 4 on page 4).
2. Pipette the samples into the small wells. You can simultaneously measure a maximum of 16 or 32
samples when using the μDrop or μDrop Duo Plate, respectively.
3. Carefully close the lid.
4. Check that each well contains an evenly distributed sample and that there are no visible air bubbles
or superfluous particles present.
5. Tilt the μDrop and μDrop Duo Plates shortly upwards to remove possibly remaining air bubbles.
Cleaning
Clean the low-volume area after each measurement. A dry silicon-free lens tissue or soft cloth can be
used to wipe down the samples.
For cleaning instructions, refer to “Cleaning the μDrop and μDrop Duo Plates” on page 19.
Cleaning verification
The cleanliness of the μDrop or μDrop Duo Plate can be verified by a simple measurement using
distilled deionized laboratory water.
1. Pipette at least 2 μl of distilled deionized laboratory water into each well. Check that there are no
air bubbles in the sample.
2. Measure the absorbance in each well with the daily routine measurement wavelength.

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Cleaning
Thermo Fisher Scientific Thermo Scientific μDrop and μDrop Duo Plates User Manual 11
3. The absorbance value of a well-cleaned sample position should be below 0.050 Abs with
wavelengths above 260 nm. If the absorbance level is too high, clean the μDrop or μDrop Duo
Plate thoroughly. For further instructions, refer to “Cleaning the μDrop and μDrop Duo Plates”
on page 19.
A spectral measurement is also a suitable tool to verify, for example, traces of DNA in the sample
positions.

2Low-volume measurements with SkanIt Software
Cleaning
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Cuvette measurements with SkanIt Software
The cuvette slot of the μDrop or μDrop Duo Plate can be used to perform photometric measurements
from standard cuvettes.
The μDrop and μDrop Duo Plates are compatible with standard rectangular cuvettes with stopper:
• Dimensions: 12.5 (W) x 12.5 (D) x 47–54 (H) mm
• Beam center height: 8.5 mm
• Beam window: ≥ 4 mm
Figure 11. Examples of an acceptable (1.) and unacceptable (2.) stopper.
Placing the cuvette
The cuvette is placed into the cuvette slot with the stopper pointing towards the adjustment screw and
the reading window facing upwards (see Figure 12).
Note Use only cuvettes with stoppers, for example, Hellma® cuvettes, Article No. 114B-10-40 or
115B-10-40.
Note Use only quartz cuvettes, black wall cuvettes are recommended for accuracy and precision.
Note Make sure that the cuvette is full. If there is air inside the cuvette, the air will move to the top
when the cuvette is horizontal and distort the measurement results.

3Cuvette measurements with SkanIt Software
Creating a new measurement session
14 Thermo Scientific μDrop and μDrop Duo Plates User Manual Thermo Fisher Scientific
Figure 12. Cuvette inserted (μDrop Plate), showing the adjustment screw (1).
The screw should be adjusted according to the length of the cuvette with stopper. You do not need to
tighten the screw completely. This will allow convenient handling of the cuvettes in measurement
series.
Because of the measurement direction, possible air bubbles present in the liquid can cause severe errors
to the results. It is particularly important to follow good pipetting practices. The reverse pipetting
technique is strongly recommended.
1. Pipette the sample into the cuvette in a sufficient volume.
2. Close the stopper and check that there are no visible air bubbles or superfluous particles present in
the sample.
3. Place the cuvette into the cuvette slot and tighten the adjustment screw, see Figure 12. Ensure that
there are no air bubbles in the measurement location (white spot, see Figure 1 on page 2).
Creating a new measurement session
User-made sessions
To create user-made measurement sessions for cuvette measurement, follow the normal SkanIt
Software procedure. However, there are certain special features to observe:
1. Plate Layout
Use the Thermo Scientific μDrop Plate or Thermo Scientific μDrop Duo Plate plate template in
SkanIt Software. Select the correct template from the Change plate template dropdown menu.
Conduct the cuvette measurement from position G11 of the μDrop or μDrop Duo Plate.