Balanced steady-state free precession (bSSFP) sequences have received increased interest for functional BOLD experiments since, in contrast to conventional EPI-based techniques, they are not prone to geometric distortions in the phase encoding direction. Characterization of measured BOLD signal changes requires consideration of extravascular as well as intravascular contributions. Knowledge about the relaxation rates of human blood is a prerequisite to quantify the intravascular contribution to the BOLD effect. Here, R1 and R2 relaxation rates of blood samples are intrinsically obtained from a series of phase-cycled bSSFP scans to account for the repetition time dependence of R2 due to rapid refocusing.
Blood sample preparation.
Fresh vein blood (20-30ml) was obtained from a healthy adult volunteer. A blood sample of ~10ml was centrifuged in order to separate plasma from RBCs. The extracted plasma was used to lower the Hct level. The oxygenation level was increased by bubbling air through the blood and gently mixing it. The Y and Hct values were assessed quantitatively using a radiometer after heating the blood samples in a water bath to a temperature of 37°C. In total, four blood samples were prepared in 3ml tubes; two of them with oxygenation levels reflecting resting state (~75%, referred to as deoxygenated) and the other two reflecting activated state (~95%, referred to as oxygenated). The two blood samples of each Y level differed in their Hct values (cf. Table 1).
MR acquisition at 3T.
The prepared blood samples were placed in a cylindrical water phantom, connected to a temperature-controlled water pump to maintain a temperature of 37°C (switched off during the MR scans). The phantom was attached to an MR-compatible robot-arm, enabling to rotate the blood samples inbetween subsequent scans (maximal duration/scan: 7min) and thus to avoid RBC sedimentation. 3D phase-cycled bSSFP experiments were performed for 20 RF phase increments ϕ in the range (0°,360°) at a resolution of 1.2x1.2x2 mm3 (matrix size: 128x64x18). Based on a Fourier transform of the measured complex bSSFP frequency profile, R1 and R2 were estimated using MIRACLE 6. The bSSFP scans were repeated for TRs ranging from 4ms to 10ms in steps of 1ms at αopt=10°. The optimal flip angle αopt in terms of SNR in the R1 and R2 maps was found based on Monte-Carlo simulations with 10000 runs by adding 1% Gaussian white noise versus R1/R2 values reported for blood at 3T 7,8 (cf. Fig. 1). Gold standard R1 and R2 were derived based on single-slice inversion recovery with variable inversion times TI=[25, 200, 1000, 2000, 4000]ms and single-slice single-echo spin-echo with variable echo times TE=[10, 30, 75, 150, 300]ms. Gold standard R2* was obtained from a multi-echo gradient-echo sequence with echo times ranging from 4.92ms to 245ms.
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