/* ghn_cacb.c 3D HNCACB gradient sensitivity enhanced version. Correlates Ca(i) and Cb(i) with N(i), NH(i), and N(i+1), NH(i+1). Uses constant time evolution for the 15N shifts. Standard features include maintaining the 13C carrier in the Cab region throughout using off-res SLP pulses; square pulses on Cab with first null at 13CO; one lobe sinc pulses on 13CO with first null at Ca; one lobe sinc pulse to put H2O back along z (the sinc one-lobe is significantly more selective than gaussian, square, or seduce 90 pulses); optional 2H decoupling when CaCb magnetization is transverse for 4 channel spectrometers. Magic-angle option for coherence transfer gradients. TROSY option for N15/H1 evolution/detection. pulse sequence: Wittekind and Mueller, JMR B101, 201 (1993) Muhandiram and Kay, JMR, B103, 203 (1994) Kay, Xu, and Yamazaki, JMR, A109, 129-133 (1994) SLP pulses: J Magn. Reson. 96, 94-102 (1992) TROSY: Weigelt, JACS, 120, 10778 (1998) Modified from hnco_3c_pfg_laue.c by RM 12/11/92 to add gradient SE. Modified by LEK Sept. 19, 1993, Nov 26, 1993, and Dec. 22, 1993 to minimally excite water etc. Revised and improved to a standard format by MRB, BKJ and GG for the BioPack, January 1997, so as to include calculation of the above standard features within the pulse sequence code and associated macro. TROSY added Dec 98, based on similar addition to gNhsqc. (Version Dec 1998). Semi-Constant Time option - Eriks Kupce, Oxford, 26.01.2006, based on example provided by Marco Tonelli at NMRFAM CHOICE OF DECOUPLING AND 2D MODES Set dm = 'nnn', dmm = 'ccc' Set dm2 = 'nny', dmm2 = 'ccg' (or 'ccw', or 'ccp') for 15N decoupling. Set dm3 = 'nnn' for no 2H decoupling, or 'nyn' and dmm3 = 'cwc' for 2H decoupling. Made the waltz16 field strength enterable (waltzB1) in Hz. (GG jan03) Must set = 1,2 and phase2 = 1,2 for States-TPPI acquisition in t1 [13C] and t2 [15N]. The flag f1180/f2180 should be set to 'y' if t1/t2 is to be started at halfdwell time. This will give 90, -180 phasing in f1/f2. If it is set to 'n' the phasing should be 0,0 and will still give a perfect baseline. Thus, set f1180='n' for (0,0) in 13C and f2180='n' for (0,0) in 15N. f1180='y' is ignored if ni=0, and f2180='y' is ignored if ni2=0. DETAILED INSTRUCTIONS FOR USE OF ghn_cacb 1. Obtain a printout of the Philosopy behind the BioPack development, and General Instructions using the macro: "printon man('BioPack') printoff". These Detailed Instructions for ghn_cacb may be printed using: "printon man('ghn_cacb') printoff". 2. Apply the setup macro "ghn_cacb". This loads the relevant parameter set and also sets ni=ni2=0 and phase=phase2=1 ready for a 1D spectral check. At the middle of the t1 period, the 180 degree pulses on CO and 15N are swapped to a 180 degree pulse on Ca, for the first increment of t1, to refocus Ca chemical-shift evolution ensuring a zero first-order phase correction in F1. This is also the case for the 1D spectral check, or for 2D/15N spectra, when ni=0. 3. The delay tauCC is set to 0.6 ms for 1D spectra or for 2D/15N spectra to provide a large positive signal. The normal value for 2D/13C or 3D nmr is 0.0035 (3.5 ms) which yields Ca and Cb signals of opposite sign. These approximately cancel in the 1D spectrum (see JMR B101, 201 for an explanation). tauCC is automatically reset to the value in the dg2 parameter set (normally 3.5 ms) for 2D/13C and 3D work (ie when ni>1). 4. Centre H1 frequency on H2O (4.7ppm), C13 frequency on 46ppm, and N15 frequency on the amide region (120 ppm). The C13 frequency remains at 46ppm, ie at CaCb throughout the sequence. 5. The normal 13C 180 degree pulse on CO at the middle of t1 induces a phase shift, which should be field-invariant, and so this phase shift has been calibrated and compensated in the pulse sequence. This phase shift can be checked by setting ni=1 whereby a special 1D method is invoked in which both the 13C CO 180 degree pulse and the simultaneous 15N 180 degree pulse are applied just as for all t1 times other than t1=0. First eliminate the CO pulse by setting pwC7=0 and obtain a 1D spectrum. This spectrum will have reduced intensity compared to ni=0 because of 13Ca chemical-shift evolution during the time of the 180 pulses. If the phase shift is adequately compensated, a second very similar 1D spectrum will be obtained with pwC7=pwC7a. Unlike ghn_co and ghn_ca, the most sensitive comparison of the two spectra with pwC7=0,pwC7a is obtained with phase=1 rather than phase=2. If not adequately compensated, the first increment will be out of phase with all succeeding increments and a zero-order phase-shift will be necessary in F1, which is easily done after the 2D/3D transform. Alternatively, change the calibration by changing the phshift7 parameter in the INITIALIZE VARIABLES section of the code. The pulse pwC7 is automatically reset to its calibrated value (=pwC7a) within the pulse sequence code for 3D work and 2D/t1 studies. DO NOT CHANGE pwC7a from its calibrated value. 6. H2O preservation is achieved according to Kay et al, except that a sinc one-lobe selective pulse is used to put H2O back along z. This is much more selective than a hard, Seduce1, or gaussian pulse. 7. Another difference from the work of Kay et al is that the phases of the first and last Cab 90 degree pulses are alternated to eliminate artifacts from the CO 180 degree pulse. 8. tauCC (3.5 ms) and timeTN (12.5 ms, determined for alphalytic protease) and are listed in dg2 for possible readjustment by the user. 9. TROSY: Set TROSY='y' and dm2='nnn' for a TROSY spectrum of the bottom right peak of the 2D coupled NH quartet (high-field H1, low-field N15). The TROSY spectrum gives 50% S/N compared to the decoupled spectrum for a small peptide. To select any of the other three peaks of the 2D coupled quartet, in a clockwise direction from bottom right, change t4/t10 from x/y to x/-y to -x/-y to -x/y. NOTE, the phases of the SE train are almost the same as those determined for the gNhsqc sequence. The major difference is that kappa is eliminated compared to normal ghn_cacb so the N15 magnetization has not evolved with respect to the attached H's. I.e. the N15 state would be Ix rather than IySz if there was no coherence gradient - this imparts a 90 degree shift so t8 is changed to y (from x in the normal spectrum). Also gzlvl1 is after the 180 N15 pulse rather than before as in gNhsqc, so the sign of icosel and the t4/t10 phase2 increments are also swapped compared to gNhsqc. For ghn_c... type sequences, H1 decoupling during the first timeTN is replaced by a pi pulse at kappa/2 to reduce S/N loss for large molecules during the first TN period. For these sequences H2O flipback is achieved with two sinc one-lobe pulses, an additional one just before the SE train, similar to gNhsqc. 12. PROJECTION-RECONSTRUCTION experiments: Projection-Reconstruction experiments are enabled by setting the projection angle, pra to values between 0 and 90 degrees (0 < pra < 90). Note, that for these experiments axis='ph', ni>1, ni2=0, phase=1,2 and phase2=1,2 must be used. Processing: use wft2dx macro for positive tilt angles and wft2dy for negative tilt angles. wft2dx = wft2d(1,0,-1,0,0,1,0,1,0,1,0,1,-1,0,1,0) wft2dy = wft2d(1,0,-1,0,0,-1,0,-1,0,1,0,1,1,0,-1,0) The following relationships can be used to inter-convert the frequencies (in Hz) between the tilted, F1(+)F3, F1(-)F3 and the orthogonal, F1F3, F2F3 planes: F1(+) = F1*cos(pra) + F2*sin(pra) F1(-) = F1*cos(pra) - F2*sin(pra) F1 = 0.5*[F1(+) + F1(-)]/cos(pra) F2 = 0.5*[F1(+) - F1(-)]/sin(pra) References: E.Kupce and R.Freeman, J. Amer. Chem. Soc., vol. 125, pp. 13958-13959 (2003). E.Kupce and R.Freeman, J. Amer. Chem. Soc., vol. 126, pp. 6429-6440 (2004). Related: S.Kim and T.Szyperski, J. Amer. Chem. Soc., vol. 125, pp. 1385-1393 (2003). Eriks Kupce, Oxford, 26.08.2004. */ #include static int /* T is for TROSY='y', phx etc also enable TROSY phase changes */ phx[1]={0}, phy[1]={1}, phi3[2] = {0,2}, phi5[4] = {0,0,2,2}, phi9[8] = {0,0,0,0,2,2,2,2}, rec[4] = {0,2,2,0}, recT[4] = {3,1,1,3}; static double d2_init=0.0, d3_init=0.0; pulsesequence() { /* DECLARE AND LOAD VARIABLES */ char f1180[MAXSTR], /* Flag to start t1 @ halfdwell */ f2180[MAXSTR], /* Flag to start t2 @ halfdwell */ SCT[MAXSTR], /* Semi-constant time flag for N-15 evolution */ mag_flg[MAXSTR], /* magic-angle coherence transfer gradients */ TROSY[MAXSTR]; /* do TROSY on N15 and H1 */ int icosel, /* used to get n and p type */ t1_counter, /* used for states tppi in t1 */ t2_counter, /* used for states tppi in t2 */ PRexp, /* projection-reconstruction flag */ ni2 = getval("ni2"); double tau1, /* t1 delay */ tau2, /* t2 delay */ tauCC = getval("tauCC"), /* delay for Ca to Cb cosy */ timeTN = getval("timeTN"), /* constant time for 15N evolution */ t2a=0.0, t2b=0.0, halfT2=0.0, CTdelay=0.0, kappa = 5.4e-3, lambda = 2.4e-3, csa, sna, pra = M_PI*getval("pra")/180.0, pwClvl = getval("pwClvl"), /* coarse power for C13 pulse */ pwC = getval("pwC"), /* C13 90 degree pulse length at pwClvl */ rf0, /* maximum fine power when using pwC pulses */ /* 90 degree pulse at Cab (46ppm), first off-resonance null at CO (174ppm) */ pwC1, /* 90 degree pulse length on C13 at rf1 */ rf1, /* fine power for 5.1 kHz rf for 600MHz magnet */ /* 180 degree pulse at Ca (46ppm), first off-resonance null at CO(174ppm) */ pwC2, /* 180 degree pulse length at rf2 */ rf2, /* fine power for 11.4 kHz rf for 600MHz magnet */ /* the following pulse lengths for SLP pulses are automatically calculated */ /* by the macro "proteincal". SLP pulse shapes, "offC7" etc are called */ /* directly from your shapelib. */ pwC7 = getval("pwC7"), /*180 degree pulse at CO(174ppm) null at Ca(56ppm) */ pwC7a = getval("pwC7a"), /* pwC7a=pwC7, but not set to zero when pwC7=0 */ phshift7, /* phase shift induced on Cab by pwC7 ("offC7") pulse */ pwZ, /* the largest of pwC7 and 2.0*pwN */ pwZ1, /* the larger of pwC7a and 2.0*pwN for 1D experiments */ rf7, /* fine power for the pwC7 ("offC7") pulse */ compH = getval("compH"), /* adjustment for C13 amplifier compression */ compC = getval("compC"), /* adjustment for C13 amplifier compression */ pwHs = getval("pwHs"), /* H1 90 degree pulse length at tpwrs */ tpwrs, /* power for the pwHs ("H2Osinc") pulse */ waltzB1 = getval("waltzB1"), /* waltz16 field strength (in Hz) */ pwHd, /* H1 90 degree pulse length at tpwrd */ tpwrd, /* 7.5 kHz rf for WALTZ decoupling */ pwNlvl = getval("pwNlvl"), /* power for N15 pulses */ pwN = getval("pwN"), /* N15 90 degree pulse length at pwNlvl */ sw1 = getval("sw1"), sw2 = getval("sw2"), gt1 = getval("gt1"), /* coherence pathway gradients */ gzcal = getval("gzcal"), /* g/cm to DAC conversion factor */ gzlvl1 = getval("gzlvl1"), gzlvl2 = getval("gzlvl2"), gt0 = getval("gt0"), /* other gradients */ gt3 = getval("gt3"), gt4 = getval("gt4"), gt5 = getval("gt5"), gt7 = getval("gt7"), gstab = getval("gstab"), gzlvl0 = getval("gzlvl0"), gzlvl3 = getval("gzlvl3"), gzlvl4 = getval("gzlvl4"), gzlvl5 = getval("gzlvl5"), gzlvl6 = getval("gzlvl6"), gzlvl7 = getval("gzlvl7"), gzlvl8 = getval("gzlvl8"); getstr("f1180",f1180); getstr("f2180",f2180); getstr("SCT",SCT); getstr("mag_flg",mag_flg); getstr("TROSY",TROSY); /* LOAD PHASE TABLE */ settable(t2,1,phy); settable(t3,2,phi3); settable(t4,1,phx); settable(t5,4,phi5); if (TROSY[A]=='y') {settable(t8,1,phy); settable(t9,1,phx); settable(t10,1,phy); settable(t11,1,phx); settable(t12,4,recT);} else {settable(t8,1,phx); settable(t9,8,phi9); settable(t10,1,phx); settable(t11,1,phy); settable(t12,4,rec);} /* INITIALIZE VARIABLES */ if( dpwrf < 4095 ) { printf("reset dpwrf=4095 and recalibrate C13 90 degree pulse"); psg_abort(1); } /* set tauCC to 0.6ms for 1D spectral check, otherwise it will be the */ /* value in the dg2 parameter set (about 3.5ms) for 2D/13C and 3D work */ if (ni>1) tauCC = tauCC; else tauCC = 0.0006; /* maximum fine power for pwC pulses */ rf0 = 4095.0; /* 90 degree pulse on Cab, null at CO 128ppm away */ pwC1 = sqrt(15.0)/(4.0*128.0*dfrq); rf1 = (compC*4095.0*pwC)/pwC1; rf1 = (int) (rf1 + 0.5); /* 180 degree pulse on Cab, null at CO 128ppm away */ pwC2 = sqrt(3.0)/(2.0*128.0*dfrq); rf2 = (4095.0*compC*pwC*2.0)/pwC2; rf2 = (int) (rf2 + 0.5); if( rf2 > 4095 ) { printf("increase pwClvl so that C13 90 < 22us*(600/sfrq)"); psg_abort(1);} /* 180 degree one-lobe sinc pulse on CO, null at Ca 118ppm away */ rf7 = (compC*4095.0*pwC*2.0*1.65)/pwC7a; /* needs 1.65 times more */ rf7 = (int) (rf7 + 0.5); /* power than a square pulse */ /* the pwC7 pulse at the middle of t1 */ if ((ni2 > 0.0) && (ni == 1.0)) ni = 0.0; if (pwC7a > 2.0*pwN) pwZ = pwC7a; else pwZ = 2.0*pwN; if ((pwC7==0.0) && (pwC7a>2.0*pwN)) pwZ1=pwC7a-2.0*pwN; else pwZ1=0.0; if ( ni > 1 ) pwC7 = pwC7a; if ( pwC7 > 0 ) phshift7 = 320.0; else phshift7 = 0.0; /* selective H20 one-lobe sinc pulse */ tpwrs = tpwr - 20.0*log10(pwHs/(compH*pw*1.69)); /* needs 1.69 times more */ tpwrs = (int) (tpwrs); /* power than a square pulse */ /* power level and pulse time for WALTZ 1H decoupling */ pwHd = 1/(4.0 * waltzB1) ; /* 7.5 kHz rf */ tpwrd = tpwr - 20.0*log10(pwHd/(compH*pw)); tpwrd = (int) (tpwrd + 0.5); /* set up Projection-Reconstruction experiment */ tau1 = d2; tau2 = d3; PRexp=0; csa = 1.0; sna = 0.0; if((pra > 0.0) && (pra < 90.0)) /* PR experiments */ { PRexp = 1; csa = cos(pra); sna = sin(pra); tau1 = d2*csa; tau2 = d2*sna; } /* CHECK VALIDITY OF PARAMETER RANGES */ if(SCT[A] == 'n') { if (PRexp) { if( 0.5*ni*sna/sw1 > timeTN - WFG3_START_DELAY) { printf(" ni is too big. Make ni equal to %d or less.\n", ((int)((timeTN - WFG3_START_DELAY)*2.0*sw1/sna))); psg_abort(1);} } else { if ( 0.5*ni2*1/(sw2) > timeTN - WFG3_START_DELAY) { printf(" ni2 is too big. Make ni2 equal to %d or less.\n", ((int)((timeTN - WFG3_START_DELAY)*2.0*sw2))); psg_abort(1);} } } if( tauCC < (gt7 + 1.0e-4)) tauCC = (gt7 + 1.0e-4); if ( dm[A] == 'y' || dm[B] == 'y' || dm[C] == 'y' ) { printf("incorrect dec1 decoupler flags! Should be 'nnn' "); psg_abort(1);} if ( dm2[A] == 'y' || dm2[B] == 'y' ) { printf("incorrect dec2 decoupler flags! Should be 'nny' "); psg_abort(1);} if ( dm3[A] == 'y' || dm3[C] == 'y' ) { printf("incorrect dec3 decoupler flags! Should be 'nyn' or 'nnn' "); psg_abort(1);} if ( dpwr2 > 50 ) { printf("dpwr2 too large! recheck value "); psg_abort(1);} if ( pw > 20.0e-6 ) { printf(" pw too long ! recheck value "); psg_abort(1);} if ( pwN > 100.0e-6 ) { printf(" pwN too long! recheck value "); psg_abort(1);} if ( TROSY[A]=='y' && dm2[C] == 'y') { text_error("Choose either TROSY='n' or dm2='n' ! "); psg_abort(1);} /* PHASES AND INCREMENTED TIMES */ /* Phase incrementation for hypercomplex 2D data, States-Haberkorn element */ if (phase1 == 2) { tsadd(t3,3,4); tsadd(t2,3,4);} if (TROSY[A]=='y') { if (phase2 == 2) icosel = +1; else {tsadd(t4,2,4); tsadd(t10,2,4); icosel = -1;} } else { if (phase2 == 2) {tsadd(t10,2,4); icosel = +1;} else icosel = -1; } /* Calculate modifications to phases for States-TPPI acquisition */ if( ix == 1) d2_init = d2; t1_counter = (int) ( (d2-d2_init)*sw1 + 0.5 ); if(t1_counter % 2) { tsadd(t3,2,4); tsadd(t12,2,4); } if( ix == 1) d3_init = d3; t2_counter = (int) ( (d3-d3_init)*sw2 + 0.5 ); if(t2_counter % 2) { tsadd(t8,2,4); tsadd(t12,2,4); } /* Set up CONSTANT/SEMI-CONSTANT time evolution in N15 */ halfT2 = 0.0; CTdelay = timeTN + pwC2 + WFG_START_DELAY - SAPS_DELAY; if(ni>1) { if(f1180[A] == 'y') /* Set up f1180 */ tau1 += 0.5*csa/sw1; /* if not PRexp then csa = 1.0 */ if(PRexp) { halfT2 = 0.5*(ni-1)/sw1; /* ni2 is not defined */ if(f1180[A] == 'y') { tau2 += 0.5*sna/sw1; halfT2 += 0.25*sna/sw1; } t2b = t1_counter*((halfT2 - CTdelay)/(ni-1)); } } if (ni2>1) { halfT2 = 0.5*(ni2-1)/sw2; if(f2180[A] == 'y') /* Set up f2180 */ { tau2 += 0.5/sw2; halfT2 += 0.25/sw2; } t2b = t2_counter*((halfT2 - CTdelay)/(ni2-1)); } tau1 = tau1/2.0; tau2 = tau2/2.0; if(tau1 < 0.2e-6) tau1 = 0.0; if(tau2 < 0.2e-6) tau2 = 0.0; if(t2b < 0.0) t2b = 0.0; t2a = CTdelay - tau2 + t2b; if(t2a < 0.2e-6) t2a = 0.0; /* uncomment these lines to check t2a and t2b printf("%d: t2a = %.12f", t2_counter,t2a); printf(" ; t2b = %.12f\n", t2b); */ /* BEGIN PULSE SEQUENCE */ status(A); delay(d1); if ( dm3[B] == 'y' ) { lk_hold(); lk_sampling_off();} /*freezes z0 correction, stops lock pulsing*/ rcvroff(); obspower(tpwr); decpower(pwClvl); dec2power(pwNlvl); decpwrf(rf0); obsoffset(tof); txphase(zero); delay(1.0e-5); if (TROSY[A] == 'n') dec2rgpulse(pwN, zero, 0.0, 0.0); /*destroy N15 and C13 magnetization*/ decrgpulse(pwC, zero, 0.0, 0.0); zgradpulse(gzlvl0, 0.5e-3); delay(1.0e-4); if (TROSY[A] == 'n') dec2rgpulse(pwN, one, 0.0, 0.0); decrgpulse(pwC, zero, 0.0, 0.0); zgradpulse(0.7*gzlvl0, 0.5e-3); delay(5.0e-4); rgpulse(pw,zero,0.0,0.0); /* 1H pulse excitation */ dec2phase(zero); zgradpulse(gzlvl0, gt0); delay(lambda - gt0); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); txphase(one); zgradpulse(gzlvl0, gt0); delay(lambda - gt0); rgpulse(pw, one, 0.0, 0.0); obspower(tpwrs); if (TROSY[A]=='y') {txphase(two); shaped_pulse("H2Osinc", pwHs, two, 5.0e-4, 0.0); obspower(tpwr); zgradpulse(gzlvl3, gt3); delay(2.0e-4); dec2rgpulse(pwN, zero, 0.0, 0.0); delay(0.5*kappa - 2.0*pw); rgpulse(2.0*pw, two, 0.0, 0.0); obspower(tpwrd); /* POWER_DELAY */ decphase(zero); dec2phase(zero); decpwrf(rf2); delay(timeTN - 0.5*kappa - POWER_DELAY); } else {txphase(zero); shaped_pulse("H2Osinc", pwHs, zero, 5.0e-4, 0.0); obspower(tpwrd); zgradpulse(gzlvl3, gt3); delay(2.0e-4); dec2rgpulse(pwN, zero, 0.0, 0.0); txphase(one); delay(kappa - pwHd - 2.0e-6 - PRG_START_DELAY); rgpulse(pwHd,one,0.0,0.0); txphase(zero); delay(2.0e-6); obsprgon("waltz16", pwHd, 90.0); /* PRG_START_DELAY */ xmtron(); decphase(zero); dec2phase(zero); decpwrf(rf2); delay(timeTN - kappa); } sim3pulse(0.0, pwC2, 2.0*pwN, zero, zero, zero, 0.0, 0.0); decphase(t3); decpwrf(rf1); delay(timeTN); dec2rgpulse(pwN, zero, 0.0, 0.0); if (TROSY[A]=='n') {xmtroff(); obsprgoff(); rgpulse(pwHd,three,2.0e-6,0.0);} zgradpulse(gzlvl3, gt3); delay(2.0e-4); if ( dm3[B] == 'y' ) /* begins optional 2H decoupling */ { gt7=0.0; /* no gradients during 2H decoupling */ dec3rgpulse(1/dmf3,one,10.0e-6,2.0e-6); dec3unblank(); dec3phase(zero); delay(2.0e-6); setstatus(DEC3ch, TRUE, 'w', FALSE, dmf3); } decrgpulse(pwC1, t3, 0.0,0.0); zgradpulse(gzlvl7, gt7); decpwrf(rf2); decphase(zero); delay(tauCC - gt7); decrgpulse(pwC2, zero, 0.0, 0.0); zgradpulse(gzlvl7, gt7); decpwrf(rf1); decphase(t2); txphase(one); delay(tauCC - gt7 - pwHd - 2.0e-6 - PRG_START_DELAY); rgpulse(pwHd,one,0.0,0.0); txphase(zero); delay(2.0e-6); obsprgon("waltz16", pwHd, 90.0); /* PRG_START_DELAY */ xmtron(); decrgpulse(pwC1, t2, 0.0, 0.0); decphase(zero); /* xxxxxxxxxxxxxxxxxxxxxx 13Cab EVOLUTION xxxxxxxxxxxxxxxxxx */ if ((ni>1.0) && (tau1>0.0)) /* total 13C evolution equals d2 exactly */ { /* 2.0*pwC1/PI compensates for evolution at 64% rate during pwC1 */ decpwrf(rf7); if(tau1 - 2.0*pwC1/PI - WFG3_START_DELAY - 0.5*pwZ > 0.0) { delay(tau1 - 2.0*pwC1/PI - WFG3_START_DELAY - 0.5*pwZ); /* WFG3_START_DELAY */ sim3shaped_pulse("", "offC7", "", 0.0, pwC7a, 2.0*pwN, zero, zero, zero, 0.0, 0.0); initval(phshift7, v7); decstepsize(1.0); dcplrphase(v7); /* SAPS_DELAY */ delay(tau1 - 2.0*pwC1/PI - SAPS_DELAY - 0.5*pwZ - 2.0e-6); } else { initval(180.0, v7); decstepsize(1.0); dcplrphase(v7); /* SAPS_DELAY */ delay(2.0*tau1 - 4.0*pwC1/PI - SAPS_DELAY - 2.0e-6); } } else if (ni==1.0) /* special 1D check of pwC7 phase enabled when ni=1 */ { decpwrf(rf7); delay(10.0e-6 + SAPS_DELAY + 0.5*pwZ1 + WFG_START_DELAY); /* WFG3_START_DELAY */ sim3shaped_pulse("", "offC7", "", 0.0, pwC7, 2.0*pwN, zero, zero, zero, 2.0e-6, 0.0); initval(phshift7, v7); decstepsize(1.0); dcplrphase(v7); /* SAPS_DELAY */ delay(10.0e-6 + WFG3_START_DELAY + 0.5*pwZ1); } else /* 13Ca evolution refocused for 1st increment */ { decpwrf(rf2); decrgpulse(pwC2, zero, 2.0e-6, 0.0); } decphase(one); decpwrf(rf1); decrgpulse(pwC1, one, 2.0e-6, 0.0); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); zgradpulse(gzlvl8, gt7); decpwrf(rf2); decphase(zero); delay(tauCC - gt7 - pwHd - 2.0e-6 - PRG_STOP_DELAY); decrgpulse(pwC2, zero, 0.0, 0.0); zgradpulse(gzlvl8, gt7); decpwrf(rf1); decphase(t5); delay(tauCC - gt7); decrgpulse(pwC1, t5, 0.0, 0.0); /* xxxxxxxxxxxxxxxxxx OPTIONS FOR N15 EVOLUTION xxxxxxxxxxxxxxxxxxxxx */ dec2phase(t8); txphase(one); dcplrphase(zero); if ( dm3[B] == 'y' ) /* turns off 2H decoupling */ { setstatus(DEC3ch, FALSE, 'c', FALSE, dmf3); dec3rgpulse(1/dmf3,three,2.0e-6,2.0e-6); dec3blank(); lk_autotrig(); /* resumes lock pulsing */ } zgradpulse(gzlvl4, gt4); decphase(zero); decpwrf(rf2); delay(2.0e-4); if (TROSY[A]=='n') {rgpulse(pwHd,one,0.0,0.0); txphase(zero); delay(2.0e-6); obsprgon("waltz16", pwHd, 90.0); xmtron();} dec2rgpulse(pwN, t8, 0.0, 0.0); /* N15 EVOLUTION BEGINS HERE */ dec2phase(t9); if(SCT[A] == 'y') { delay(t2a); dec2rgpulse(2.0*pwN, t9, 0.0, 0.0); delay(t2b); decrgpulse(pwC2, zero, 0.0, 0.0); /* WFG_START_DELAY */ } else { delay(timeTN - tau2); sim3pulse(0.0, pwC2, 2.0*pwN, zero, zero, t9, 0.0, 0.0); } dec2phase(t10); decpwrf(rf7); if (TROSY[A]=='y') { if (tau2 > gt1 + 2.0*GRADIENT_DELAY + 1.5e-4 + pwHs) { txphase(three); delay(timeTN - pwC7a - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(tau2 - gt1 - 2.0*GRADIENT_DELAY - 1.5e-4 - pwHs); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwrs); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); shaped_pulse("H2Osinc", pwHs, three, 0.0, 0.0); txphase(t4); obspower(tpwr); /* POWER_DELAY */ delay(0.5e-4 - POWER_DELAY); } else if (tau2 > pwHs + 0.5e-4) { txphase(three); delay(timeTN-pwC7a-WFG_START_DELAY-gt1-2.0*GRADIENT_DELAY-1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwrs); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(tau2 - pwHs - 0.5e-4); shaped_pulse("H2Osinc", pwHs, three, 0.0, 0.0); txphase(t4); obspower(tpwr); /* POWER_DELAY */ delay(0.5e-4 - POWER_DELAY); } else { txphase(three); delay(timeTN - pwC7a - WFG_START_DELAY - gt1 - 2.0*GRADIENT_DELAY - 1.5e-4 - pwHs); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwrs); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); /* WFG_START_DELAY */ shaped_pulse("H2Osinc", pwHs, three, 0.0, 0.0); txphase(t4); obspower(tpwr); /* POWER_DELAY */ delay(0.5e-4 - POWER_DELAY); decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(tau2); } } else { if (tau2 > kappa) { delay(timeTN - pwC7a - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); delay(kappa - gt1 - 2.0*GRADIENT_DELAY - 1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); } else if (tau2 > (kappa - pwC7a - WFG_START_DELAY)) { delay(timeTN + tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); /* WFG_START_DELAY */ decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(kappa -pwC7a -WFG_START_DELAY -gt1 -2.0*GRADIENT_DELAY -1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); } else if (tau2 > gt1 + 2.0*GRADIENT_DELAY + 1.0e-4) { delay(timeTN + tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); delay(kappa - tau2 - pwC7a - WFG_START_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(tau2 - gt1 - 2.0*GRADIENT_DELAY - 1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); } else { delay(timeTN + tau2 - kappa - PRG_STOP_DELAY - pwHd - 2.0e-6); xmtroff(); obsprgoff(); /* PRG_STOP_DELAY */ rgpulse(pwHd,three,2.0e-6,0.0); txphase(t4); delay(kappa-tau2-pwC7a-WFG_START_DELAY-gt1-2.0*GRADIENT_DELAY-1.0e-4); if (mag_flg[A]=='y') magradpulse(gzcal*gzlvl1, gt1); else zgradpulse(gzlvl1, gt1); /* 2.0*GRADIENT_DELAY */ obspower(tpwr); /* POWER_DELAY */ delay(1.0e-4 - POWER_DELAY); /* WFG_START_DELAY */ decshaped_pulse("offC7", pwC7a, zero, 0.0, 0.0); delay(tau2); } } /* xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx */ if (TROSY[A]=='y') rgpulse(pw, t4, 0.0, 0.0); else sim3pulse(pw, 0.0, pwN, t4, zero, t10, 0.0, 0.0); txphase(zero); dec2phase(zero); zgradpulse(gzlvl5, gt5); if (TROSY[A]=='y') delay(lambda - 0.65*(pw + pwN) - gt5); else delay(lambda - 1.3*pwN - gt5); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); zgradpulse(gzlvl5, gt5); txphase(one); dec2phase(t11); delay(lambda - 1.3*pwN - gt5); sim3pulse(pw, 0.0, pwN, one, zero, t11, 0.0, 0.0); txphase(zero); dec2phase(zero); zgradpulse(gzlvl6, gt5); delay(lambda - 1.3*pwN - gt5); sim3pulse(2.0*pw, 0.0, 2.0*pwN, zero, zero, zero, 0.0, 0.0); dec2phase(t10); zgradpulse(gzlvl6, gt5); if (TROSY[A]=='y') delay(lambda - 1.6*pwN - gt5); else delay(lambda - 0.65*pwN - gt5); if (TROSY[A]=='y') dec2rgpulse(pwN, t10, 0.0, 0.0); else rgpulse(pw, zero, 0.0, 0.0); delay((gt1/10.0) + 1.0e-4 +gstab - 0.5*pw + 2.0*GRADIENT_DELAY + POWER_DELAY); rgpulse(2.0*pw, zero, 0.0, rof1); dec2power(dpwr2); /* POWER_DELAY */ if (mag_flg[A] == 'y') magradpulse(icosel*gzcal*gzlvl2, gt1/10.0); else zgradpulse(icosel*gzlvl2, gt1/10.0); /* 2.0*GRADIENT_DELAY */ delay(gstab); rcvron(); statusdelay(C,1.0e-4 - rof1); if (dm3[B]=='y') lk_sample(); setreceiver(t12); }