Friday, April 15, 2016

Whole Blood Collection Card for Protein Microarray

Whole blood collection cards to acquire and stabilize, RNA, DNA and Protein from lancet "finger-stick" collection methods are important tools in life sciences and healthcare.  The best and most sensitive assays require sample input of the highest integrity.  The old saying is especially true in the laboratory, "Garage in garbage out".  The sample input integrity is essential to acquiring great results.

The most important aspect of any dry blood spot collection card is the purity of the collection matrix that the blood sample is applied.  There can be no contamination on the matrix that will compromise the sample being collected. All material used to manufacture the collection matrix should use the highest purity materials validated by chromatography and use high purity deionized water and chemicals in the process. The final collection matrix holding the whole blood must be free of naturally occurring RNases, DNases, and Proteases, the enzymes that break down and degrade these important biomolecules that exist everywhere! Therefore, the cards should be assembled in clean room environments free of contamination.  Each collection matrix must be homogenous in composition.  Uniformity, thickness, and absorbency characteristics must be consistent across all cards.  

The collection matrix used for Arrayit Whole Blood Collection Cards are enclosed in a convenient cover with a flap that easily opens and closes to protect the blood spot collection area from environmental contamination or damage.  Each is bar coded for unique identification and has space on the front of the card for information the specimen collection personnel may want write on the card.  Logos and text can all be customized to meet customers specific requirements. Cards can come in bulk or packed individually in environmental proof bags.

For use in protein microarray, cards come with specially formulated elution/reaction buffers with spin columns to prepare the sample to be applied to protein microarrays.  Here is a training video of the process of elution.

In general, a single 1/2 inch circle is enough for a single robust protein microarray reaction.  There is no need to separate serum and internal Arrayit studies have shown that whole blood eluted from the card in our reaction buffer can provide superior results to wet serum alone in specific applications. The process of separating serum from whole blood can selectively eliminate or reduce the concentration of certain proteins and other biomolecules that may be desirable to measure on a microarray.  One 1/2 circle of eluate mixed with the reaction buffer provided with the kit yields a 120 microliter reaction cocktail ready to apply to a microarray.  For larger microarrays, more circles may be used.  For lower volume microarray reactions, partial use of the cocktail may be applied to the microarray. For best results use all complimentary microarray surface chemistry, buffers, and solutions.  Traditional microarray and ELISA buffers are not suitable for proper elution and reaction.  Contact me for more details, 

Thursday, March 24, 2016

High Throughput Blood Testing

How we run the equivalent of 10,368 blood based ELISA assays on only 4 microscope slide size pieces of glass in a single tool with the footprint of a standard 96 well plate using blood input levels from fingerstick collection. I realize this looks complicated and perhaps even sounds impossible to you. But it's all quite simple really and it works. Tracking the data generated on our miniaturized multiplexed and parallelized microarray testing platform is critical. The first photo is the form we use in the lab for our AHC4x12 hybridization cassette (also pictured). The slide numbers, block numbers for microarray quantification, and blood sample numbers are all fixed based on the cassette hardware and the microarray manufacturing pattern which are displayed on the form. Each blood sample is reacted to 2 microarray "blocks" of spots for up to 48 samples per tool (see 2 photos of single blocks here). We write on this form the slide number, date, microarray type, and bar code number from the blood sample that is applied to the microarray. It works and tracks everything perfect. Total test time only 4 hours all the way through to final data reports, 1 technician, total sample preparation and applying the sample to the microarrays requires only 144 pipetting steps for 10,368 data points.
Form Used in the lab to track blood samples
AHC4x12 Hybridization Cassette.  Each "well" is a microarray of 2 blocks of spots.
 Example block 1
Example Block 2
(16-bit raw image data presented in pseudo color scale)

Friday, December 4, 2015

2015 SpotBot Microarrayer Publications

Sometimes customers ask us if we have any references for our SpotBot Microarrayer.  Here is a list of the publications published in 2015.

A Manrakhan, PR Stephen, PJR Cronje - Crop Protection, 2015 - Elsevier
The phytotoxicity of the fruit fly bait GF-120 NF (containing 0.24 g/L spinosad and fruit fly
attractants including a protein hydrolysate, ammonium acetate and.
A Mendoza, DM Torrisi, S Sell, NC Cady, DA Lawrence - Analyst, 2016 -
... Anti-CD marker and heat shock protein antibodies were initially diluted in PBS buffer (pH 7.4)
to working concentration, 0.5 mg mL −1 , transferred to a 384-well microtiter plate (Thermofisher,
IL), and immediately placed in the Arrayit robotic microarray spotter, 
SpotBot II (Arrayit ...
S Roy, JH Soh, JY Ying - Biosensors and Bioelectronics, 2016 - Elsevier
... Microarrays were printed on functionalized glass slides using the SpotBot® 3 Personal
Microarrayers (Sunnyvale, CA). 2.2. 
... 10 μM of aminated hairpin or linear CPs was then dissolved
in 5× SSC (pH 7), and spotted in a microarray format using the 
SpotBot 3 microarrayer. ...
MM Jørgensen, R Bæk, K Varming - Journal of extracellular …, 2015 -
... Contact printing. Microarray printing was performed on a SpotBot ® Extreme Protein
Edition Microarray Printer with a 946MP4 pin (ArrayIt, CA, USA). Temperature and
humidity were kept at 15–18°C and 55–65%, respectively. 
KR Jakobsen, BS Paulsen, R Bæk… - Journal of …, 2015 -
... Phenotyping of extracellular vesicles using the EV Array. Production of microarrays.
Microarray printing was performed on a 
SpotBot ® Extreme Protein Edition Microarray
Printer with a 946MP4 pin (ArrayIt Corporation, CA, USA). 
Y Tsai, J Cutts, A Kimura, D Varun, DA Brafman - Stem cell research, 2015 - Elsevier
... A contact arrayer (SpotBot® 2, Arrayit®) was used to print the polymers. The printing conditions
were a 1000 ms inking time and a 250 ms stamping time. Each spot had a diameter of 150–200
μm and neighboring spots were separated by a center-to-center distance of 450 μm. 
D PatelA HaqueY GaoA Revzin - Integrative Biology, 2015 -
... 30 min at room temperature prior to printing. Protein microarrays were contact-printed
under ambient conditions on silane-modified 75 × 25 mm 2 glass slides using a
spotbot arrayer. Pins collected protein from a 384-well plate ...
Y Blanco, M Moreno-Paz, J AguirreV Parro - Springer
... Equipment. Spectrophotometer, ultrasonicator, microcentrifuge, robot arrayer for multiple
slides (eg MicroGrid II TAS Arrayer from Digilab and 
SpotBot ® 3 Personal Microarrayers
from Arrayit), scanner for fluorescence (see step f of Sect. 
S Gündisch, L Annaratone, C Beese, E Drecol… - Laboratory …, 2015 -
... Analysis of Protein Expression by Reverse Phase Protein Arrays (RPPA). RPPAs
were generated by using the 
SpotBot Extreme microarrayer according to the
manufacturer's instructions (Arrayit, Sunnyvale, CA 94089, USA). 
M Šunderić, A Šedivá, D Robajac… - Biotechnology and …, 2015 - Wiley Online Library
... The spotting was performed using SpotBot ® 3 Personal Microarrayer Protein Edition
(Arrayit Corporation, Sunnyvale, CA) at the temperature of 14 °C and humidity of 60%.
The samples were spotted into 14 identical arrays on the slide. 
K Kaur, NY Zheng, K Smith, M Huang, L Li, NT Pauli… - 2015 -
... Antibody Microarray. Antibodies were diluted in protein printing buffer to 250ug/ml and printed
on SuperEpoxy glass slides using the 
SpotBot 3 microarrayer (ArrayIt). Antibodies were spotted
in triplicate on each array. Printed slides were stored overnight at 4°C in the dark. 
F Zhang, A Briones, M Soloviev - Peptide Microarrays: Methods and …, 2015 - Springer
... For the methods described here, a smaller scale desktop microarray instruments would be the
most suitable. For example 
SpotBot ® 2 Personal Microarrayer from Arrayit corporation which
is capable of printing up to 384 samples onto the maximum of 14 slides. 3. 
R Yentrapalli, O Azimzadeh, A Kraemer… - Journal of …, 2015 - Elsevier
... 2.6.3. RPPA analysis. RPPA analysis was performed as previously described [28].
RPPAs were generated using the 
SpotBot Extreme Microarray Spotter according
to manufacturer's instructions (Anopoli, Eichgraben, Austria). 
S Boellner, KF Becker - Microarrays, 2015 -
... Automated systems that are commonly used for RPPA spotting are ArrayIt SpotBot Extreme
Microarray Spotter [62] (Arrayit, Sunnyvale, CA, USA), Aushon BioSystems 2470 Microarrayer
[19] (Billerica, MA, USA), or SpotArray Microarray printing system [63] (Perkin Elmer 
R Ramji, NT Khan, A Muñoz-Rojas, K Miller-Jensen - RSC Advances, 2015 -
... All images were processed using ImageJ software. Protein spotting and cell culture.
A commercial microarray spotter, 
SpotBot 3 (Arrayit Inc., USA) was used to spot
FITC-BSA inside the PDMS microwell array patterned on the glass slide. 
Y DengA Ediriwickrema, F Yang, J Lewis, M Girardi… - Nature materials, 2015 -
A water-resistant sunblock based on bioadhesive nanoparticles encapsulating
a model ultraviolet filter at low concentrations adheres to the stratum corneum
without subsequent intra-epidermal or follicular penetration.
CJH Dunand, PE Leon, K Kaur, GS Tan… - The Journal of …, 2015 - Am Soc Clin Investig
... 18). Antibody microarray. Eighty-three H3N2-reactive antibodies were diluted in
Protein Printing Buffer (ArrayIt) to 250 μg/ml and printed in triplicate on SuperEpoxy
glass slides using the 
SpotBot 3 microarrayer (ArrayIt). Before ...
M ThielenT Speck, R Seidel - Royal Society …, 2015 -
Skip to main content. ...
Y Lu, Q Xue, MR Eisele, ES Sulistijo… - Proceedings of the …, 2015 - National Acad Sciences
N Kumar, J Richter, J Cutts, KT Bush, C Trujillo… - eLife, 2015 -
An expandable cell population derived from human pluripotent stem cells exhibits properties
of mesoderm and is restricted to differentiate into derivatives of intermediate mesoderm.
M Waespy, TT Gbem, L Elenschneider, AP Jeck… - PLoS Negl Trop …, 2015 -
Author Summary In this study we demonstrated the binding of TconTS lectin domains
(TconTS-LD) to high-mannose N -glycans and provide evidence for a biological function for this
interaction. TconTS1 and TconTS2 lectin domain bind to galactosyl as well as mannosyl glycans 
C Shirley - English Literary Renaissance, 2015 - Wiley Online Library
... The first quoted above is a complaint directed to an apparently virtuous beloved of indeterminate
gender: to men that knows ye not. ye may aper to be. ffol cher and with owt 
spotbot sewarly un
to me. so ys yowr wonted kynd. be proffe so sewarly knowen. that I wel not be blynd. 
R Fan, Y Lu, J Chen - US Patent App. 14/629,164, 2015 - Google Patents
The present invention relates to a system, device, and method for the high throughput multiplexed
detection of a wide number of compounds. The invention comprises of a microwell array coupled
to a capture agent array to form a plurality of interfaces between a microwell and a 
Page 1. POLITECNICO DI MILANO Scuola di Ingegneria Industriale e dell'Informazione CORSO
soft-litografiche per la realizzazione di patterns biomolecolari su superfici antifouling di