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Functional Connectivity

using iRSFC toolbox
Sunghyon Kyeong
Institute of Behavioural Science in Medicine, 

Yonsei University College of Medicine
iRSFC
intuitive resting-state functional connectivity
(iRSFC) toolbox
2
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Step 1 - Dataset and Directory
3
Subject List

피험자 리스트가 기록되어 있는 엑셀
데이터를 선택.
헤더가 subject 인 컬럼에 폴더명에
해당되는 피험자 이름을 기록.
DATA path

데이터의 위치를 지정한다. 

데이터는 다음과 같이 구조로 

저장되어 있어야 함.
(DATA path)/NOR001/rest
(DATA path)/NOR002/rest
OUT path

분석결과 및 ROI mask가 

저장될 위치를 지정.
<Subject List 엑셀 파일 예시>
iRSFC intuitive resting-state functional connectivity (iRSFC) toolbox
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Step 2 - Filter & Regressors
4
Prefix of preprocessed images

SPM8을 이용하여 전처리를 했다면 spatial preprocessing이 완료된
뇌영상 데이터에는 원래 뇌영상 데이터 이름 앞에 'sw' 또는 'swa' 가
prefix로 붙음.
Regressors

resting state fMRI 데티어의
시계열 분석에서 confounding effects가 될
가능성이 있는 noise signal들을 제거해주는
역할.
Global Signal을 제거하느냐? 마느냐? 에 대
해서는 의견이 분분함.
Dummy Scan

자기장이 안정화
될때까지의 데이터를 분석에서
제외함. 초기 8~12s 정도.
TR [s]

fMRI 촬영에
사용된 TR을
초 단위로 입력
Bandpass Filter [Hz]

fMRI 촬영에 사용된
TR을 초 단위로 입력
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Step 3 - Select ROIs
5
: Both atlas based and user defined ROIs can be used for the seed-based FC analysis.
(1a)
(2a) (2b)
(1b)
(3)
Make ROI mask

sphere or box 모양의 ROI를 생성할 수 있음.

(MNI or Talairach coordinate 좌표 모두 사용 가능함.)
(1b): (1a)에서 생성한 이미지 형태의 ROI를 선택.

(1b)와 (2b)를 동시에 선택하는 것이 가능함.
AAL or Dosenbach
with Ctrl-key, 

select multiple ROIs
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Create ROIs
6
(1a)
Make ROI mask

sphere or box 모양의 ROI를 생성할 수 있음.

(MNI or Talairach coordinate 좌표 모두 사용 가능함.)
go to slide 19 page
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Step 4 - Run Analysis
7
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Now, we have z-maps
8
PCC L NAcc R DLPFC
for each seed ROI and each subject
9
with covariates
Two-sample t-test
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Specify 2nd-level
10
Group 1 scans

Group 1의 connectivity maps을 선택
Group 2 scans

Group 2의 connectivity maps을 선택
Covariates

covariate으로 입력할 변수의 갯수만큼

New:Covariate으로 선택
Design

어떤 통계분석을 진행할지 선택함

choose Two-sample t-test
Directory

결과 파일이 저장될 위치를 지정
Vector

Group1 subjects의 covariates을 sequential 하게 입
력하고 바로 뒤이어 Group2 subjects의 covariates을
입력
Name

Covariate의 이름 입력
27 25 29 43 39 41 57 24 29 24
ex) Age in Group 1, in Group 2
모든 정보가 입력 되었으면 “▶Run Batch”를 클
릭하여 프로그램을 실행한다.
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Estimation of Parameters
11
Select SPM.mat

Slide의 8페이지에서 지정했던 Directory에 생성된
SPM.mat 파일을 선택함.
Specify 2nd-level 을 통해서
구성한 General Linear Model의 parameter를 추정(계산)함.
▶Run Batch를 클릭하여 프로그램을 실행.
▶
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Contrast manager
12
(1)
(2)
(3a)
(4a)
1 -1 0 0
Group1>Group2 contrast는 Group1에서
증가된 FC or Group2에서 감소된 FC를 의미함.
(4b) -1 1 0 0
(3b) Group1 < Group2
Group1 > Group2 Contrast(s)
(5)
Age Sex
Correlation analysis 

between FC and clinical scales
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p 14
in Results
(1)
(2)
(3)
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p 15
Save Results as MASK
To extract connectivity value in
each cluster, regional clusters
should be saved as mask
image.
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Extract FC using iRSFC
16
page 14에서 저장했던 mask image를
선택함. 또는 Maks ROI mask를 통해
서 직접 그린 ROI를 선택할 수도 있음.
추출하고 싶은 데이터가 위치한 경로를
입력함. 데이터 구조는 

다음과 같이 이루어져 있어야 함.
(Data path)Lt_Amygdalasubj001.img
(Data path)Lt_Amygdalasubj002.img
(Data path)Lt_Amygdalasubj003.img
(Data path)Lt_Amygdalasubj004.img
결과 파일이 저장될 폴더를 선택함.
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Example of extracted FC
17
filename cl01 cl02 cl03 cl04
zscore_Talairach_-12_8_-8_sphere_3mm_CON1 -0.8 -1.3 -0.61 -0.84
zscore_Talairach_-12_8_-8_sphere_3mm_CON2 -2.48 -3.64 -1.13 -0.79
zscore_Talairach_-12_8_-8_sphere_3mm_CON3 -1.1 -1.6 0.36 -0.34
zscore_Talairach_-12_8_-8_sphere_3mm_CON4 -0.66 -2.83 -2.14 -1.6
zscore_Talairach_-12_8_-8_sphere_3mm_CON5 -1.05 -2.2 -1.24 -1.81
zscore_Talairach_-12_8_-8_sphere_3mm_PAT1 1.36 0.82 0.93 2.09
zscore_Talairach_-12_8_-8_sphere_3mm_PAT2 3.53 2.07 1.37 1.01
zscore_Talairach_-12_8_-8_sphere_3mm_PAT3 -0.79 0.23 1.55 1.4
zscore_Talairach_-12_8_-8_sphere_3mm_PAT4 1.51 0.25 2.2 2.45
zscore_Talairach_-12_8_-8_sphere_3mm_PAT5 1.85 0.28 5.19 2.75
open tables.csv file which was saved in (Output path)
18
ROI connectivity Analysis
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Create ROIs
19
(1)
Make ROI mask

sphere or box 모양의 ROI를 생성할 수 있음.

(MNI or Talairach coordinate 좌표 모두 사용 가능함.)
Region of Interest x y z
Left TPJ -50 -31 23
Left ACC -7 21 36
Left Broca’s Area -41 12 14
Left Amygdala -24 -5 -18
Left Insula -36 8 4
Right TPJ 50 -31 23
Right ACC 7 21 36
Right Homotopy of Broca’s Area 41 12 14
Right Amygdala 24 -5 -18
Right Insula 36 8 4
(2)
(3)
(4)
(5)
(6)
(7)
Reference for ROIs, A. Vercammen et al. BIOL PSYCHIATRY 2010 67:912-918, http://neuro.imm.dtu.dk/services/jerne/ninf/voi.html
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Step 4 - Run Analysis
20
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
Check OUTPUT
21
node number ROI file name
1 Lt_ACC
2 Lt_Amyg
3 Lt_Broca
4 Lt_Insula
5 Lt_TPJ
6 Rt_ACC
7 Rt_Amyg
8 Rt_Broca
9 Rt_Insula
10 Rt_TPJ
subjname ROI_1-2 ROI_1-3 ROI_1-4 ROI_1-5 ROI_1-6 ROI_1-7
CON1-20130825-KSK 0.068 0.408 0.818 0.123 0.957 -0.048
CON2-20130825-KKM -0.164 0.577 0.62 0.051 0.753 -0.05
CON3-20130901-KMY -0.28 0.081 0.202 -0.006 0.684 -0.392
CON4-20130929-KSB 0.02 0.146 0.739 0.095 0.371 -0.131
CON5-20131006-HTH -0.245 0.039 0.1 0.154 0.203 -0.156
SPR1-20130901-CCS 0.058 0.229 0.514 0.301 0.577 -0.006
SPR2-20130901-OKS 0.218 -0.236 -0.262 0.128 0.264 -0.187
SPR3-20130901-BY -0.017 0.033 0.021 -0.215 0.342 -0.081
SPR4-20131013-SSH 0.069 0.115 0.372 0.207 0.625 0.131
SPR5-20130929-YMJ 0.225 0.275 0.663 0.046 0.624 0.01
Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p
References for Atlas
• AAL Atlas (n=116)

N. Tzourio-Mazoyer et al. (2002), Automated Anatomical Labeling of Activations in SPM Using a
Macroscopic Anatomical Parcellation of the MNI MRI Single-Subject Brain, NeuroImage 15,
273-289 (2002)
• Dosenbach Atlas (n=160)

Nico U. F. Dosenbach et al. (2010), Prediction of Individual Brain Maturity Using fMRI, Science
329:5997 pp.1358-1361.

n=160, sphere shape, radius =5 mm, minimum distances between ROI center = 10 mm, no
overlap among ROIs
22

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Intuitive Resting-State Functional Connectivity (iRSFC) Toolbox

  • 1. Functional Connectivity
 using iRSFC toolbox Sunghyon Kyeong Institute of Behavioural Science in Medicine, 
 Yonsei University College of Medicine
  • 2. iRSFC intuitive resting-state functional connectivity (iRSFC) toolbox 2
  • 3. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Step 1 - Dataset and Directory 3 Subject List
 피험자 리스트가 기록되어 있는 엑셀 데이터를 선택. 헤더가 subject 인 컬럼에 폴더명에 해당되는 피험자 이름을 기록. DATA path
 데이터의 위치를 지정한다. 
 데이터는 다음과 같이 구조로 
 저장되어 있어야 함. (DATA path)/NOR001/rest (DATA path)/NOR002/rest OUT path
 분석결과 및 ROI mask가 
 저장될 위치를 지정. <Subject List 엑셀 파일 예시> iRSFC intuitive resting-state functional connectivity (iRSFC) toolbox
  • 4. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Step 2 - Filter & Regressors 4 Prefix of preprocessed images
 SPM8을 이용하여 전처리를 했다면 spatial preprocessing이 완료된 뇌영상 데이터에는 원래 뇌영상 데이터 이름 앞에 'sw' 또는 'swa' 가 prefix로 붙음. Regressors
 resting state fMRI 데티어의 시계열 분석에서 confounding effects가 될 가능성이 있는 noise signal들을 제거해주는 역할. Global Signal을 제거하느냐? 마느냐? 에 대 해서는 의견이 분분함. Dummy Scan
 자기장이 안정화 될때까지의 데이터를 분석에서 제외함. 초기 8~12s 정도. TR [s]
 fMRI 촬영에 사용된 TR을 초 단위로 입력 Bandpass Filter [Hz]
 fMRI 촬영에 사용된 TR을 초 단위로 입력
  • 5. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Step 3 - Select ROIs 5 : Both atlas based and user defined ROIs can be used for the seed-based FC analysis. (1a) (2a) (2b) (1b) (3) Make ROI mask
 sphere or box 모양의 ROI를 생성할 수 있음.
 (MNI or Talairach coordinate 좌표 모두 사용 가능함.) (1b): (1a)에서 생성한 이미지 형태의 ROI를 선택.
 (1b)와 (2b)를 동시에 선택하는 것이 가능함. AAL or Dosenbach with Ctrl-key, 
 select multiple ROIs
  • 6. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Create ROIs 6 (1a) Make ROI mask
 sphere or box 모양의 ROI를 생성할 수 있음.
 (MNI or Talairach coordinate 좌표 모두 사용 가능함.) go to slide 19 page
  • 7. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Step 4 - Run Analysis 7
  • 8. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Now, we have z-maps 8 PCC L NAcc R DLPFC for each seed ROI and each subject
  • 10. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Specify 2nd-level 10 Group 1 scans
 Group 1의 connectivity maps을 선택 Group 2 scans
 Group 2의 connectivity maps을 선택 Covariates
 covariate으로 입력할 변수의 갯수만큼
 New:Covariate으로 선택 Design
 어떤 통계분석을 진행할지 선택함
 choose Two-sample t-test Directory
 결과 파일이 저장될 위치를 지정 Vector
 Group1 subjects의 covariates을 sequential 하게 입 력하고 바로 뒤이어 Group2 subjects의 covariates을 입력 Name
 Covariate의 이름 입력 27 25 29 43 39 41 57 24 29 24 ex) Age in Group 1, in Group 2 모든 정보가 입력 되었으면 “▶Run Batch”를 클 릭하여 프로그램을 실행한다.
  • 11. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Estimation of Parameters 11 Select SPM.mat
 Slide의 8페이지에서 지정했던 Directory에 생성된 SPM.mat 파일을 선택함. Specify 2nd-level 을 통해서 구성한 General Linear Model의 parameter를 추정(계산)함. ▶Run Batch를 클릭하여 프로그램을 실행. ▶
  • 12. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Contrast manager 12 (1) (2) (3a) (4a) 1 -1 0 0 Group1>Group2 contrast는 Group1에서 증가된 FC or Group2에서 감소된 FC를 의미함. (4b) -1 1 0 0 (3b) Group1 < Group2 Group1 > Group2 Contrast(s) (5) Age Sex
  • 13. Correlation analysis 
 between FC and clinical scales
  • 14. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p 14 in Results (1) (2) (3)
  • 15. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p 15 Save Results as MASK To extract connectivity value in each cluster, regional clusters should be saved as mask image.
  • 16. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Extract FC using iRSFC 16 page 14에서 저장했던 mask image를 선택함. 또는 Maks ROI mask를 통해 서 직접 그린 ROI를 선택할 수도 있음. 추출하고 싶은 데이터가 위치한 경로를 입력함. 데이터 구조는 
 다음과 같이 이루어져 있어야 함. (Data path)Lt_Amygdalasubj001.img (Data path)Lt_Amygdalasubj002.img (Data path)Lt_Amygdalasubj003.img (Data path)Lt_Amygdalasubj004.img 결과 파일이 저장될 폴더를 선택함.
  • 17. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Example of extracted FC 17 filename cl01 cl02 cl03 cl04 zscore_Talairach_-12_8_-8_sphere_3mm_CON1 -0.8 -1.3 -0.61 -0.84 zscore_Talairach_-12_8_-8_sphere_3mm_CON2 -2.48 -3.64 -1.13 -0.79 zscore_Talairach_-12_8_-8_sphere_3mm_CON3 -1.1 -1.6 0.36 -0.34 zscore_Talairach_-12_8_-8_sphere_3mm_CON4 -0.66 -2.83 -2.14 -1.6 zscore_Talairach_-12_8_-8_sphere_3mm_CON5 -1.05 -2.2 -1.24 -1.81 zscore_Talairach_-12_8_-8_sphere_3mm_PAT1 1.36 0.82 0.93 2.09 zscore_Talairach_-12_8_-8_sphere_3mm_PAT2 3.53 2.07 1.37 1.01 zscore_Talairach_-12_8_-8_sphere_3mm_PAT3 -0.79 0.23 1.55 1.4 zscore_Talairach_-12_8_-8_sphere_3mm_PAT4 1.51 0.25 2.2 2.45 zscore_Talairach_-12_8_-8_sphere_3mm_PAT5 1.85 0.28 5.19 2.75 open tables.csv file which was saved in (Output path)
  • 19. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Create ROIs 19 (1) Make ROI mask
 sphere or box 모양의 ROI를 생성할 수 있음.
 (MNI or Talairach coordinate 좌표 모두 사용 가능함.) Region of Interest x y z Left TPJ -50 -31 23 Left ACC -7 21 36 Left Broca’s Area -41 12 14 Left Amygdala -24 -5 -18 Left Insula -36 8 4 Right TPJ 50 -31 23 Right ACC 7 21 36 Right Homotopy of Broca’s Area 41 12 14 Right Amygdala 24 -5 -18 Right Insula 36 8 4 (2) (3) (4) (5) (6) (7) Reference for ROIs, A. Vercammen et al. BIOL PSYCHIATRY 2010 67:912-918, http://neuro.imm.dtu.dk/services/jerne/ninf/voi.html
  • 20. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Step 4 - Run Analysis 20
  • 21. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p Check OUTPUT 21 node number ROI file name 1 Lt_ACC 2 Lt_Amyg 3 Lt_Broca 4 Lt_Insula 5 Lt_TPJ 6 Rt_ACC 7 Rt_Amyg 8 Rt_Broca 9 Rt_Insula 10 Rt_TPJ subjname ROI_1-2 ROI_1-3 ROI_1-4 ROI_1-5 ROI_1-6 ROI_1-7 CON1-20130825-KSK 0.068 0.408 0.818 0.123 0.957 -0.048 CON2-20130825-KKM -0.164 0.577 0.62 0.051 0.753 -0.05 CON3-20130901-KMY -0.28 0.081 0.202 -0.006 0.684 -0.392 CON4-20130929-KSB 0.02 0.146 0.739 0.095 0.371 -0.131 CON5-20131006-HTH -0.245 0.039 0.1 0.154 0.203 -0.156 SPR1-20130901-CCS 0.058 0.229 0.514 0.301 0.577 -0.006 SPR2-20130901-OKS 0.218 -0.236 -0.262 0.128 0.264 -0.187 SPR3-20130901-BY -0.017 0.033 0.021 -0.215 0.342 -0.081 SPR4-20131013-SSH 0.069 0.115 0.372 0.207 0.625 0.131 SPR5-20130929-YMJ 0.225 0.275 0.663 0.046 0.624 0.01
  • 22. Sunghyon Kyeong (Yonsei University) intuitive Resting State Functional Connectivity (iRSFC) toolbox p References for Atlas • AAL Atlas (n=116)
 N. Tzourio-Mazoyer et al. (2002), Automated Anatomical Labeling of Activations in SPM Using a Macroscopic Anatomical Parcellation of the MNI MRI Single-Subject Brain, NeuroImage 15, 273-289 (2002) • Dosenbach Atlas (n=160)
 Nico U. F. Dosenbach et al. (2010), Prediction of Individual Brain Maturity Using fMRI, Science 329:5997 pp.1358-1361.
 n=160, sphere shape, radius =5 mm, minimum distances between ROI center = 10 mm, no overlap among ROIs 22