Wednesday, May 29, 2013

Protocol Separate Serum from Whole Blood without a Centrifuge

Protocol: Separate Serum from Whole Blood without a Centrifuge and only a finger stick amount of whole blood.

fingerstick
Arrayit Blood Card fingerstick lancet. Blood sampling is achieved by placing the middle finger on your left hand directly on the red, spring-loaded sampling platform as shown and pressing down firmly. Follow instructions carefully before, during and after use.
http://www.arrayit.com/Microarray_Diagnostics/Blood_Cards/blood_cards.html
blood card collection
Arrayit Blood Card shown 10 minutes after placing 5 drops of whole human blood inside the red box. The red blood cell components remain on the left portion of the linear transfer membrane and the serum (yellowish fraction) partitions to the right. Arrayit Blood Cards can be used for DNA, mRNA, serum and protein analysis by taking sample discs of these respective fractions.
Blood Sampling (Short Protocol)1. Wash your hands thoroughly with soap and hot water for 1 minute.
2. Open the plastic product envelop and place the kit components on the benchtop.
3. Remove the plastic safety cap from the lancet.
4. Place the lancet on the bench top with the red sampling platform facing upward.
5. Place your left middle fingertip on the lancet sampling platform.
6. Press down firmly on the lancet.
7. Place 5 large drops of whole blood inside the red box on the Blood Card.
8. Allow the blood to separate on the Blood Card for 5 minutes.
9. Allow the Blood Card to dry for 5 minutes.
10. Place the Blood Card in the silver anti-static shipping envelop for storage and shipping.

http://www.arrayit.com/Microarray_Diagnostics/Blood_Cards/blood_cards.html
Blood Sampling (Complete Protocol)1. Wash your hands thoroughly with soap and hot water for 1 minute. This step is important because it removes bacteria and other contaminants from your hands prior to sampling. The use of hot water also improves blood circulation in your finger tips, which improves the efficiency of the sampling procedure. Make sure to wash for at least 1 minute as shorter wash periods will reduce blood flow. Please avoid topical cleaning with alcohol swabs and other solvents as this will dehydrate the sampling area and reduce blood flow.
2. Open the plastic product envelop and place the kit components on the benchtop. Use a scissors or other cutting implement to carefully cut the top of the plastic envelop and then remove the Blood Card, Customer Reference Card, two lancets and the silver anti-static shipping envelope. The kit components (see Figure 4) should be placed on a smooth, clean and level surface such as a laboratory bench or clinical table.
3. Remove the plastic safety cap from the lancet. The pink lancets contain a clear plastic safety cap on the top of the lancet to protect the lancet from premature use. Remove the plastic cap carefully immediately prior to use.
4. Place the lancet on the laboratory bench with the red sampling platform facing upward. The lancet contains a red, spring-loaded sample platform. Make sure the sampling platform is facing upwards prior to sampling.
5. Place your left middle fingertip on the lancet sampling platform. Place the tip of your middle finger softly in the center of the sampling platform with your finger tip aligned directly in the center of the 3 mm orifice located in the center of the platform (see Figure 5). Because of the proximity to your heart, blood pressure and blood flow in the left hand is slightly greater than in your right hand, and the left middle finger is chosen for this reason.
6. Press down firmly on the lancet. Press down firmly on the lancet platform to initiate the sampling process. The lancet uses a spring-loaded stainless steel sampling device approximately 350 µm in diameter to create a nearly invisible sampling site on your left middle finger tip. The sampling process is fast and nearly undetectable.
7. Place 5 large drops of whole blood inside the red box on the Blood Card. For best results, make sure you are standing up for this step to increase blood pressure and blood flow. Use your non-sampling hand to squeeze your finger tip firmly and create five blood droplets. Place the droplets directly in the center of the red box of the Blood Card, making light contact between the droplet and the Blood Card membrane. Avoid touching the Blood Card membrane directly with your fingertip as this may interfer with the separation process. A properly lanced finger tip should produce 5 large drops or 100 µl of whole blood in less than one minute. If blood flow stops, it may be necessary to use the second lancet provided with the kit to obtain 5 large blood droplets.
8. Allow the blood to separate on the Blood Card for 5 minutes. After placing 5 large blood droplets in the center of the red box, allow 5 minutes for the blood components to separate on the Blood Card. The Blood Card should be placed level and flat on the laboratory bench during the separation process to permit even flow of the blood components. The blood components should separate from left to right on the card such that the red cell components remain to the left side of the separation membrane and the yellowish serum components migrate rightward during the 5 minute process. Five large blood droplets should produce a fractionation result similar to Figure 6. Customers using whole blood obtained by intravenous blood draws should place 5 drops or 100 µl of blood inside the red box.
9. Allow the Blood Card to dry for 5 minutes. Following the 5 minute fractionation process, incubate the Blood Card on the laboratory bench for an additional 5 minutes to allow drying of the fractionated whole blood components. The drying process enhances the stability of the blood components for sampling and storage and ensures that the components will remain firmly adhered to the Blood Card during shipping. Avoid making direct contact with the separated blood components at all times during the sampling, fractionation and drying steps as this can contaminate the samples and complicate downstream analysis.
10. Place the Blood Card in the silver anti-static shipping envelope for storage and mailing. Open the zip-lock located at the top of the silver anti-static shipping envelope and place the dried Blood Card into the envelope. Re-seal the zip-lock located at the top of the silver envelope and mail to the proper laboratory for testing. Alternatively, the Blood Card in the sealed silver anti-static envelope can be stored in a cool dark drawer or storage cabinet for future testing. Make certain to retain your Customer Reference Card as a permanent record of your sample, making sure to write your name and the sampling date on the card for future reference. The patient identification number located on your reference card is identical to the patient identification number on the Blood Card.
blood card sample
Arrayit Blood Card sample disc isolated 24 hours after whole blood collection. A 6 mm round disc containing fractionated serum was obtained using the Arrayit Blood Card Sample Disc Punch (Cat. ASDP). Serum discs of this diameter contain approximately 250 µg of total serum protein containing peptides, antigens, antibodies and other analytes of scientific, research, veterinary and pathology interest.  Arrayit Blood Card Sample Disc Punch (Cat. ASDP) enables precision manual generation of 6 mm sample discs for DNA, mRNA and serum samples. The disc punch is manufactured using injection molded plastic for resistance to light solvents. Spring loading allows easy manual operation. Sample discs are automatically released from the disc punch to allow easy transfer into microfuge tubes and microplates for elution of serum for downstream application such as protein microarray.  Email us for the serum elution protocol at arrayit@arrayit.com 
http://www.arrayit.com/Microarray_Diagnostics/Blood_Cards/blood_cards.html