How small things get big
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1 How small things get big Marcel Bruinenberg LifeLines University Medical Centre Groningen 1 Lab Automation 2009
2 Central question Why do some people get old in a healthy manner, and why have others already severe impairments or diseases early in their life? Focus on Multifactorial diseases (gene + environment) Factors which cause more than one disease
3 Area General population cohort
4 Cohort dimensions The study will cover 30,000 index persons with an age between 30 and 50 years in a defined geographical area, and subsequently assess and follow their family members father Aselect: propositus Children involved assumption: mother Parents alive assumption: partner s father partner s mother partner Family assumption: 2 children Partner assumption: child 1 child 2 Total number of participants:
5 The present situation Pilot study started November 1st, 2006 Number of locations increased to 5 currently; expected to be 12 in 2010 Over participants have been screened now. Average 70 participants per day, need to increase to >150 per day
6 Route for participants Questionnaire 1 (at home) Visit 1 Questionnaire 2 (at home) Visit 2 Repeat after 5, 10,15, 20 and 25 years
7 Content of questionnaires History of health and diseases respiratory, allergy, diabetes, heart, blood, brains, kidneys, thyroid eyes and ears, cancer Disease history of family Birth and development Lifestyle and health behavior, eating habits Quality of life Somatisation, anxiety, personality Stress and social support Person environment fit, aspirations Female health (anticonception, menstrual cycle, pregnancies)
8 Visit one Height, weight (BMI), waist and hip circumference Blood pressure: during 10 minutes Electrocardiogram (ECG) Pulmonary function (including FEV1) RFFT (Ruffs Figure Fluency Test): cognitive functioning (recognition, speed, memory) MINI: Interview on depressive complaints, fears and phobias
9 Visit two Fasting blood sample, for standard laboratory testing serum / plasma / PP plasma / DNA storage 24-hour urine collection and first morning void
10 Current Laboratory activities - Registration incoming samples - Analyzing daily routine samples - Aliquoting for Storage (-80 C; LN2) - Routine measurements (haematology, chemistry etc.) Starting: - complete automation of sample aliquoting - Isolation and Normalisation of DNA
11 LifeLines: sample challenges EDTA Heparin Citrate Serum x2 F/Ox Plastic Conical Urinalysis C Transport RT Transport 4 0 C Transport 4 0 C Transport Plasma, Buffy coat, DNA (FTA), chemistry, hormones proteomics Blood cell count, HbA1c Chemistry & Cells Serum, hormones, peptides, proteomics Glucose Urine Metabolites Bioanalytes
12 Processing of samples
13 Technical challenges Isolating DNA from 150+ participants daily High yield, long-term quality, several sample sizes, limited operator time, Illumina platform Storing 35 ml of serum, plasma, and 16 x 2 ml of urine of participants, over 5 years, in small aliquots, manageable, easily stored and retrieved, with long term quality
14 Tracking of samples 14
15 Sample storage Manual proces 15
16 Tracking and tracing Currently: in house developed software. Future: LIMS 16
17 Sample storage at present stored in Matrix 1,4 ml seprasealed cryovials For each participant EDTA1,2,3 & 4: EDTA1+2+3: EDTA4: 20 x 950 µl standard plasma 3 x 950 µl buffy coat packed cells Heparin: 3 x 1,7 ml lymphocytes # Serum Gel: 3 x 950 µl Serum 3 x 950 µl Citrate 3 x 950 µl Urine EMS 6 x 950 µl + 4 x 950 µl (AscAcid/EDTA) Urine 24H 8 x 950 µl + 4 x 950 µl (AscAcid/EDTA)
18 New building
19 New Laboratory
20 Automation platforms 150 volunteers per day gives a total of 8,100 aliquots per day Buffy Coat 450 aliquots Blood 450 aliquots 950 µl Serum 450 aliquots 950 µl Citrate 450 aliquots 950 µl Serum Gel tube 3000 aliquots 950 µl Plasma Urine aliquots EMS/EMS aliquots 24H/24H+
21 Robotics RTS- ABF 500 (retrieving plasma and buffy coats) 21
22 Robotics Tecan Freedom EVO (retrieving plasma and serum) 22
23 Robotics Synchron Urine application 23 Synchron Lymfo application (isolation of lymophocytes)
24 Robotics QIAGEN Autopure (isolation of DNA) 24
25 Robotics Synchron DNA application (processing for Autopure) 25
26 Sample storage Currently: Conventional -80 C freezer (serum, plasma, urine, DNA and buffy coats) 26 Liquid nitrogen (lymfocytes)
27 Sample storage Future: Backup in conventional -80 C freezer (serum, plasma, urine, DNA and buffy coats) Liquid nitrogen (lymfocytes) Automated sample stores 27
28 Sample storage Automated sample store at -80 C (~2 years) 28
29 Thank you for your attention time for questions 29
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