Intersection point Linear correlation equation Correlation coefficient (R) Hemocytometry Flow cytometry H.cytometry F. cytometry = 0.00 y = 2E-08x y = 2E-08x 0,9972 0,9728 ≠ 0.00 y = 2E-08x + 0,0024 y = 2E-08x + 0,0047 0,9986 0,9808 The correlation equations are then used to determine the cell number of your cell culture, where x is the cell number in…
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Flow cytometer (setups)
Running the flow cytometer BD Accuri C6 Choose between the number of events counted or velocity the cell culture is drawn through the flow cytometer Threshold – indicates which cells will be excluded from a count Wash cycles (miliQ water) – washing the nozzle Agitation cycles – agitation of the cell samples Fluorescence filters – 4 different FL1 533 ± 15…
Fluorescein
Evaluated addition of fluorescein to Synechocystis cells for standardizing 77K fluorescence spectra (excited at 435 nm). Measured absorption at peak 490nm: 0.53 stock solution was diluted 1:3 -> corrected absorption for stock solution at peak 2.12 from: https://en.wikipedia.org/wiki/Fluorescein Extinction coefficient at 490 nm = 76900 Mol-1 cm-1 Stock solution concentration: A=e*L*C C=A/(e*L) C=2.12 / (76900 cm-1 M * 1 cm…
Absorption and Fluorescence
Jablonski diagram Jablonski diagram representing energy levels and spectra. Solid arrows indicate radiative transitions as occurring by absorption (violet, blue) or emission (green for fluorescence; red for phosphorescence) of a photon. Dashed arrows represent non-radiative transitions (violet, blue, green, red). Internal conversion is a non-radiative transition, which occurs when a vibrational state of a higher electronic state is coupled to…
Absorption & Fluorescence spectra Synecho
Whole Cell Absorption 77K [wpfilebase tag=file id=330 tpl=simple /] Thylakoid and purified PSII 77K fluorescence [wpfilebase tag=file id=329 tpl=simple /] A: Excitation at 600 nm excites the phycobilisome and preferentially PSII B: Excitation at 420 nm excites the chlorophylls contained in PSI and PSII PSI , PSI-IsiA and IsiA fluorescence at 77K [wpfilebase tag=file id=328 tpl=simple /]
Sum-up of pre-experiments – Anaerobic incubation of Nannochloropsis
The experiment on anaerobic incubation of Nannochloropsis will determine what anaerobic incubation does to photosynthesis, and to the production of photosynthetic lipids, storage lipids, and ω-3 fatty acids. The pre-experiment was performed in order to find out what would be the ideal time point for making the cells anaerobic. The cells were grown in the same conditions as will be used…
Chloroplast transformation
Potential genes for deletion These genes are not essential components of PSII in Synechocystis psbX psbY psbZ These genes are involved in the light-independent chlorophyll synthesis chlL chlN [wpfilebase tag=file id=333 tpl=simple /] information concerning light-dependent and light independent chlorophyll synthesis Potential neutral region in chloroplast genome The region is downstream of atpB in the Chlamydomonas chloroplast genome (see Purton(2007)_ToolsTechChloroTrans).…
Cryopreservation of marine algae, Nannochloropsis
1 cryopreservation Cultivate N. oceanica or N. gaditana on a shaker (150 rpm, 23°C, 100 yE) to a cell concentration of 1*107 cells/ml. Place culture for 20 hours in the dark before cryopreservation. Equilibrate the CoolCell freezing container (check if black core is inside) at 4°C. Prepare a sterile filtered 20% solution of DMSO in the culture medium used to grow the algae. harvest the…
Expression of genes in the chloroplast
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