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This page provides examples for an example of the ISA-TAB-Nano Investigation files, File leveraging data from the an Nanotechnology Characterization Laboratory (NCL) as an example Investigation ( NCL200612A ). For the complete file, refer the NCL Investigation File Example.

The TABLE 1. ISA-TAB-Nano Investigation File Extensions to ISA-TAB

Section	Field	Field Status (if applicable)
INVESTIGATION	Investigation disease	Required
INVESTIGATION	Investigation disease term accession number	Required
INVESTIGATION	Investigation disease term source REF	Required
INVESTIGATION	Investigation outcome	Optional
STUDY	Study disease	Required
STUDY	Study disease term accession number	Required
STUDY	Study disease term source REF	Required
STUDY	Study outcome	Optional
STUDY ASSAYS	Study assay measurement name	Required
STUDY ASSAYS	Study assay measurement name term accession number	Required
STUDY ASSAYS	Study assay measurement name term source REF	Required

Ontology Source Reference

TABLE 2: Example Investigation File Ontology Source Reference Section

consists of the following sections:

ONTOLOGY SOURCE REFERENCE
INVESTIGATION
INVESTIGATION PUBLICATIONS
INVESTIGATION CONTACTS
MATERIAL
STUDY
- STUDY DESIGN DESCRIPTORS
- STUDY PUBLICATIONS
- STUDY FACTORS
- STUDY ASSAYS
- STUDY PROTOCOLS
- STUDY CONTACTS

The following sections provide an example of each Investigation File section.

Ontology Source Reference

Example Investigation File Ontology Source Reference Section

ONTOLOGY SOURCE REFERENCE Term Source NameMONPOUO

A	B	C	D	E	F	G
Term Source Name	MO	NPO	UO

ChEBI

PATO

NCIt

Term Source File

http://purl.bioontology.org/ontology/MO

http

Multiexcerpt include

MultiExcerptName	ExitDisclaimer
nopanel	true
PageWithExcerpt	wikicontent:Exit Disclaimer to Include

http://purl.bioontology.org/ontology/npo

Multiexcerpt include

MultiExcerptName	ExitDisclaimer
nopanel	true
PageWithExcerpt	wikicontent:Exit Disclaimer to Include

http://purl.bioontology.org/ontology/UO

Multiexcerpt include

MultiExcerptName	ExitDisclaimer
nopanel	true
PageWithExcerpt	wikicontent:Exit Disclaimer to Include

http://purl.bioontology.org/ontology/CHEBI

Multiexcerpt include

MultiExcerptName	ExitDisclaimer
nopanel	true
PageWithExcerpt	wikicontent:Exit Disclaimer to Include

http://purl.bioontology.org/ontology/PATO

Multiexcerpt include

MultiExcerptName	ExitDisclaimer
nopanel	true
PageWithExcerpt	wikicontent:Exit Disclaimer to Include

http://ncit.nci.nih.gov/

Term Source Version

1.3.1.1

2011-02-12

	v.11.11d
Term Source Description	MGED

Ontology

NanoParticle

Ontology

Unit

Ontology

Chemical

Entities of Biological Interest

Phenotype Ontology

NCI Thesaurus

...

Investigation

TABLE 3: Example Investigation File Investigation Section

INVESTIGATION

A

B
Investigation Identifier	NCL200612A
Investigation Title	Dendrimer-

Based MRI

Based MRI Contrast Agents
Investigation Description	The

goal of this investigation is to characterize a PAMAM dendrimer with an

associated gadolinium chelate MRI contrast agent.

Investigation Disease Investigation Disease Term Accession Number

Investigation

Disease Term Source REF Investigation Outcome Investigation

Submission Date	2002-11-30
Investigation

Public Release Date

2002-11-30

...


Investigation Disease
Investigation Disease Term Accession Number
Investigation Disease Term Source REF
Investigation Outcome

Investigation

...

Publications

TABLE 4: Example Investigation File Investigation Contacts Publication Section

INVESTIGATION CONTACTS

A

Investigation Person Last NameMcNeilInvestigation Person First NameScottInvestigation Person Mid InitialsEInvestigation Person Emailmcneils@mail.nih.govInvestigation Person Phone3018466939Investigation Person Fax Investigation Person AddressMSC 1050 Boyles Street, Frederick, MD 21702Investigation Person AffiliationNanotechnology Characterization LaboratoryInvestigation Person RolesinvestigatorInvestigation Person Roles Term Accession Number Investigation Person Roles Term Source REFMO

Investigation Publication

TABLE 5. Example Investigation Publication Section

...

B
Investigation PubMed ID	18095846
Investigation Publication DOI	10.2217/17435889.2.6.789
Investigation Publication Author List	Hall JB; Dobrovolskaia MA; Patri AK; McNeil SE
Investigation Publication Title	Characterization of nanoparticles for therapeutics
Investigation Publication Status	published
Investigation Publication Status Term Accession Number
Investigation Publication Status Term Source REF

Investigation Contacts

Example Investigation File Investigation Contacts Section

A	B
Investigation Person Last Name	Doe
Investigation Person First Name	John
Investigation Person Mid Initials	E
Investigation Person Email	doej@mail.nih.gov
Investigation Person Phone	1231231234
Investigation Person Fax
Investigation Person Address	Laboratory Street, City, State 111111
Investigation Person Affiliation	Doe Laboratories
Investigation Person Roles	investigator
Investigation Person Roles Term Accession Number
Investigation Person Roles Term Source REF	MO

Material

Example Investigation File Material Section

A	B
Material File Name	m_NCL-21.xls
Material Source Name	NCL-21

Study

Example Investigation File Study Section

A	B
Study Identifier

Study

TABLE 6. Example Investigation File Study Section

STUDY Study Identifier

	NCL200612A-CytoxicityLLC-PK1
Study Title

Cytotoxicity characterization

Cytotoxicity characterization in LLC-PK1 cells

Study

Submission Date2002-11-30Study Public Release Date2002-11-30Study DescriptionNanoparticle biocompatibility was

Description

Nanoparticle biocompatibility was evaluated in the porcine renal proximal tubule

cell

cell line, LLC-PK1. Cytotoxicity was determined as described in the NCL protocol

for LLC-PK1 Kidney Cytotoxicity Assay(GTA-1). Briefly, test materials were

diluted to the desired assay concentrations in cell culture media. Cells were

preincubated for 24 h prior to adding test material, reaching an approximate

confluence of 80%. Cells were exposed to test material for 6, 24 and 48 h,

and cytotoxicity was determined using the MTT cell viability and LDH

membrane integrity

membrane integrity assays.
Study Submission Date	2002-11-30
Study

Disease

Public Release Date	2002-11-30
Study Disease
Study Disease

Term Accession Number


Study

Disease Term

Disease Term Source REF


Study Outcome	NCL22,

NCL23

NCL23 and NCL24 were found to be minimally cytotoxic, under the testing conditions utilized.

Study

File Name

s_cytotoxicity-LLC-PK1.xls

Study Design Descriptors

TABLE 7. Example Investigation File Study Design Descriptors Section

A	B

STUDY DESIGN DESCRIPTORS


Study

Design Type

Design Type	comparison
Study

Design Type

Design Type Term Accession Number


Study

Design Type

Design Type Term Source REF

Study

...

Publications

TABLE 8. Example Investigation File Study Contacts Publications Section

STUDY CONTACTS

A

B
Study

Person Last NameDobrovolskaiaStudy Person First NameMarinaStudy Person Mid InitialsAStudy Person Emailmarina@mail.nih.govStudy Person Phone3018466352Study Person Fax Study Person AddressMSC 1050 Boyles Street, Frederick, MD 21702Study Person AffiliationNanotechnology Characterization LaboratoryStudy Person RolesinvestigatorStudy Person Roles Term Accession Number Study Person Roles Term Source REFMO

Study Publications

TABLE 9. Example Investigation File Study Publications Section

...

PubMed ID	18095846
Study Publication DOI	10.2217/17435889.2.6.789
Study Publication Author list	Hall JB; Dobrovolskaia MA; Patri AK; McNeil SE
Study Publication Title	Characterization of nanoparticles for therapeutics
Study Publication Status	published
Study Publication Status Term Accession Number
Study Publication Status Term Source REF

Study Factors

Example Investigation File Study Factors Section for Study Involving Biospecimens (such as in vitro, in vivo characterization). The study factor name and type must be of nanoparticle sample if the assay is applying a nanoparticle sample to a biological system.

A	B	C	D
Study Factor Name	nanoparticle sample	dose	time of exposure
Study Factor Type	nanoparticle sample	particle concentration	time of exposure
Study Factor Type Term Accession Number	NPO_1404	NPO_1830	NPO_1819
Study Factor Type Term Source REF	NPO	NPO	NPO

Example Investigation File Study Factors Section for Physico-Chemical Characterization Study. There should be no study factors of study factor type nanoparticle sample

A	B	C
Study Factor Name	temperature	solvent
Study Factor Type	temperature	solvent medium
Study Factor Type Term Accession Number	PATO_0000146	NPO_1855
Study Factor Type Term Source REF	PATO	NPO

Study Assays

Example Investigation File Study Assays Section

A	B	C
Study Assay Measurement Type	MTT Assay	LDH Release Assay
Study Assay Measurement Type Term Accession Number		NPO_1709
Study Assay Measurement Type Term Source REF		NPO
Study Assay Technology Type
Study Assay Technology Type Term Accession Number
Study Assay Technology Type Term Source REF
Study Assay Technology Platform
Study Assay Measurement Name	cell viability	LDH release
Study Assay Measurement

Study Assays

TABLE 10. Example Investigation File Study Assays Section

STUDY ASSAYS Study Assay Measurement TypeMTT AssayLDH Release AssayStudy Assay Measurement Type Term Accession Number NPO_1709Study Assay Measurement Type Term Source REF NPOStudy Assay Technology Type Study Assay Technology Type Term Accession Number Study Assay Technology Type Term Source REF Study Assay Technology Platform Study Assay Measurement Namecell viabilityLDH releaseStudy Assay Measurement

Name Term Accession Number

NPO_1343


Study

Assay Measurement

Assay Measurement Name Term Source REF

NPO


Study

Assay File

Assay File Name

a_MTT-LLCPK1.xls

a_LDH-LLCPK1.xls

Study

...

Protocols

TABLE 11. Example Investigation File Study Factors Protocols Section

STUDY FACTORS

A

B

C

Study Factor Namenanoparticle sampleparticle concentrationtime of exposureStudy Factor Name Term Accession NumberNPO_1404NPO_1830NPO_1819Study Factor Name Term Source REFNPONPONPOStudy Factor Type Study Factor Type Term Accession Number Study Factor Type Term Source REF

Study Protocols

TABLE 12. Example Investigation File Study Protocols Section

D	E	F	G	H	I	J
Study Protocol Name	Time-6-24-48 plate MTT assay	Test plate LDH assay	Time zero plate MTT assay	cell preparation in four 96-well plates	APAP positive control preparation	Triton-X100 positive control preparation	MTT assay reagent preparation	Test sample and positive control addition	LDH assay reagent preparation
Study Protocol Type				cell preparation	control preparation	control preparation	reagent preparation		reagent preparation
Study Protocol Type Term Accession Number
Study Protocol Type Term Source REF
Study Protocol Description	Test Plates: 6, 24 and 48 hour exposures (MTT Assay) 5.4.1 Remove appropriate test plate from incubator and replace media from Triton-X positive control wells (see plate format in Appendix) with 200 mL 1% Triton-X (made in Step 4.1.2). Let the plate set for 10 minutes at room temperature. Spin plate at 700 x g for 3 minutes. 5.4.2 Remove 100 mL of media from each well and transfer it to another plate, maintaining plate format. Use this plate immediately for the LDH assay (see Section 5.5). 5.4.3 Remove remaining media from original plate and discard. 5.4.4 Add 200 mL of fresh media to all wells. 5.4.5 Add 50 mL of MTT to all wells. 5.4.6 Cover in aluminum foil and incubate for 37°C for 4 hours. 5.4.7 Remove plate from incubator and spin at 700 x g for 3 minutes. 5.4.8 Remove media and MTT. 5.4.9 Add 200 mL of DMSO to each well. 5.4.10 Add 25 mL of glycine buffer to each well. Place on shaker to mix. 5.4.11 Read absorbance at 570 nm on plate reader using a reference wavelength of 680 nm.	Test Plates: 0,

STUDY PROTOCOLS Study Protocol NameTime-6-24-48 plate MTT assayTest plate LDH assayTime zero plate MTT assaycell preparation in four 96-well platesAPAP positive control preparationTriton-X-100 positive control preparationMTT assay reagent preparationTest sample and positive control additionLDH assay reagent preparationStudy Protocol Type cell preparationcontrol preparationcontrol preparationreagent preparation reagent preparationStudy Protocol Type Term Accession Number Study Protocol Type Term Source REF Study Protocol DescriptionTest Plates:

6, 24 and 48 hour exposures (

MTT Assay

LDH Assay) Adapted from Biovision LDH Cytotoxicity Assay Kit, K311-400)
5.

4.1 Remove appropriate test plate from incubator and replace media from Triton-X positive control wells (see plate format in Appendix) with 200 mL 1% Triton-X (made in Step 4.1.2). Let the plate set for 10 minutes at room temperature. Spin plate at 700 x g for 3 minutes

5.1 Add 100 mL of the Reaction Mixture (step 4.3.2) to each well of transfer plate. Shake plate on an orbital shaker briefly to mix samples.
5.

4.2 Remove 100 mL of media from each well and transfer it to another plate, maintaining plate format. Use this plate immediately for the LDH assay (see Section 5.5).

5.2 Incubate at room temperature for up to 20 minutes in the dark.
5.

5.3

Remove remaining media from original plate and discard.
    5.4.4 Add 200 mL of fresh media to all wells.
    5.4.5 Add 50 mL of MTT to all wells.
    5.4.6 Cover in aluminum foil and incubate for 37°C for 4 hours.
    5.4.7 Remove plate from incubator and spin at 700 x g for 3 minutes.
    5.4.8 Remove media and MTT.
    5.4.9 Add 200 mL of DMSO to each well.
    5.4.10 Add 25 mL of glycine buffer to each well. Place on shaker to mix.
    5.4.11 Read absorbance at 570 nm on plate reader using a reference wavelength of 680 nm.Test Plates: 0, 6, 24 and 48 hour exposures (LDH Assay)
    (Adapted from Biovision LDH Cytotoxicity Assay Kit, K311-400)
    5.5.1 Add 100 mL of the Reaction Mixture (step 4.3.2) to each well of transfer plate. Shake plate on an orbital shaker briefly to mix samples.
    5.5.2 Incubate at room temperature for up to 20 minutes in the dark.
    5.5.3 Read the plate on plate reader at 490 nm using a reference wavelength of 680 nm.5.2.1 Remove time zero plate from incubator and replace media from Triton-X positive control wells (see plate format in Appendix) with 200 mL 1% Triton-X (made in Step 4.1.2). Add 100 mL of media to the remaining wells. Let the plate set for 10 minutes at room temperature. Spin plate at 700 x g for 3 minutes.
    5.2.2 Remove 100 mL of media from each well and transfer it to another plate, maintaining plate format. Use this plate immediately for the LDH assay (see Section 5.5).
    5.2.3 Remove remaining media from original plate and discard.
    5.2.4 Add 200 mL of fresh media to all wells.
    5.2.5 Add 50 mL of MTT to all wells.
    5.2.6 Cover in aluminum foil and incubate at 37°C for 4 hours.
    5.2.7 Remove plate from incubator and spin at 700 x g for 3 minutes.
    5.2.8 Aspirate media and MTT.
    5.2.9 Add 200 mL of DMSO to all wells.
    5.2.10 Add 25 mL of glycine buffer to all wells. Place on shaker to mix.
    5.2.11 Read absorbance at 570 nm on plate reader.5.1.1 Harvest cryopreserved cells from prepared flasks (limit to 20 passages).
    5.1.2 Count cell concentration using a coulter counter or hemocytometer.
    5.1.3 Dilute cells to a density of 2.5 x 105 cells/mL in M199 (3% FBS) cell culture media.
    5.1.4 Plate 100 mL cells/well as per plate format (Appendix) for four 96-well plates (time zero, 6 hour sample exposure, 24 hour sample exposure, 48 hour sample exposure). The format indicates no cells in rows D and E as they serve as particle blanks to be subtracted from cell treatment wells. Each plate accommodates two samples (Rows A–C and F–H). Each nanoparticle is tested at nine dilutions. Column 11 receives the APAP positive control and column 12 receives Triton X-100.
    5.1.5 Incubate plates for 24 hours at 5% CO2, 37°C and 95% humidity.4.1.1 Acetaminophen (APAP) positive control: Add 19 mg to a total volume of 5 mL M199 Cell Culture Media (with 3% FBS) to make a 25 mM solution. Sterile filter using a 0.2 mm filter.4.1.2 1% Triton-X-100 positive control: Add 1 mL of Triton-X-100 to 99 mL of media. Sterile filter using a 0.2 mm filter.4.2.1 MTT solution: 5 mg/mL MTT in PBS, store for up to one month at 4°C in dark
    4.2.2 Glycine Buffer: 0.1 M glycine (MW 75.07), 0.1 M NaCl (MW 58.44),
    pH 10.5, store at room temperature.5.3.1 The highest concentration of nanoparticle tested should be at the limit of solubility. The test sample should be at physiological pH. Neutralization of acidic/basic test samples may be required.
    5.3.2 Dilute test compound in media, making a total of 9 1:4 dilutions.
    5.3.3 Add 100 mL of each sample dilution and positive control to 6 hour, 24 hour and 48 hour exposure plates as per the plate format (Appendix).4.3.1 Reconstitute catalyst in 1 mL dH20 for 10 min with occasional vortexing (stable for 2 weeks at 4°C).
    4.3.2 Reaction mixture (for one 96-well plate): Add 250 mL of reconstituted catalyst solution to 11.25 mL of dye solution (stable for 2 weeks at 4°C).Study Protocol URINCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfNCL_Method_GTA-1.pdfStudy Protocol Version1.11.11.11.11.11.11.11.11.1Study Protocol Parameters Name         Study Protocol Parameters Name Term Accession Number         Study Protocol Parameters Name Term Source REF         Study Protocol Components NameMTT; acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakeracetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; biovision LDH-cytoxicity assay kit; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakerMTT; acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shakerM199 cell culture media; fetal

Read the plate on plate reader at 490 nm using a reference wavelength of 680 nm.	5.2.1 Remove time zero plate from incubator and replace media from Triton-X positive control wells (see plate format in Appendix) with 200 mL 1% Triton-X (made in Step 4.1.2). Add 100 mL of media to the remaining wells. Let the plate set for 10 minutes at room temperature. Spin plate at 700 x g for 3 minutes. 5.2.2 Remove 100 mL of media from each well and transfer it to another plate, maintaining plate format. Use this plate immediately for the LDH assay (see Section 5.5). 5.2.3 Remove remaining media from original plate and discard. 5.2.4 Add 200 mL of fresh media to all wells. 5.2.5 Add 50 mL of MTT to all wells. 5.2.6 Cover in aluminum foil and incubate at 37°C for 4 hours. 5.2.7 Remove plate from incubator and spin at 700 x g for 3 minutes. 5.2.8 Aspirate media and MTT. 5.2.9 Add 200 mL of DMSO to all wells. 5.2.10 Add 25 mL of glycine buffer to all wells. Place on shaker to mix. 5.2.11 Read absorbance at 570 nm on plate reader.	5.1.1 Harvest cryopreserved cells from prepared flasks (limit to 20 passages). 5.1.2 Count cell concentration using a coulter counter or hemocytometer. 5.1.3 Dilute cells to a density of 2.5 x 105 cells/mL in M199 (3% FBS) cell culture media. 5.1.4 Plate 100 mL cells/well as per plate format (Appendix) for four 96-well plates (time zero, 6 hour sample exposure, 24 hour sample exposure, 48 hour sample exposure). The format indicates no cells in rows D and E as they serve as particle blanks to be subtracted from cell treatment wells. Each plate accommodates two samples (Rows A–C and F–H). Each nanoparticle is tested at nine dilutions. Column 11 receives the APAP positive control and column 12 receives Triton X-100. 5.1.5 Incubate plates for 24 hours at 5% CO2, 37°C and 95% humidity.	4.1.1 Acetaminophen (APAP) positive control: Add 19 mg to a total volume of 5 mL M199 Cell Culture Media (with 3% FBS) to make a 25 mM solution. Sterile filter using a 0.2 mm filter.	4.1.2 1% Triton-X-100 positive control: Add 1 mL of Triton-X-100 to 99 mL of media. Sterile filter using a 0.2 mm filter.	4.2.1 MTT solution: 5 mg/mL MTT in PBS, store for up to one month at 4°C in dark 4.2.2 Glycine Buffer: 0.1 M glycine (MW 75.07), 0.1 M NaCl (MW 58.44), pH 10.5, store at room temperature.	5.3.1 The highest concentration of nanoparticle tested should be at the limit of solubility. The test sample should be at physiological pH. Neutralization of acidic/basic test samples may be required. 5.3.2 Dilute test compound in media, making a total of 9 1:4 dilutions. 5.3.3 Add 100 mL of each sample dilution and positive control to 6 hour, 24 hour and 48 hour exposure plates as per the plate format (Appendix).	4.3.1 Reconstitute catalyst in 1 mL dH20 for 10 min with occasional vortexing (stable for 2 weeks at 4°C). 4.3.2 Reaction mixture (for one 96-well plate): Add 250 mL of reconstituted catalyst solution to 11.25 mL of dye solution (stable for 2 weeks at 4°C).
Study Protocol URI	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf	NCL_Method_GTA-1.pdf
Study Protocol Version	1.1	1.1	1.1	1.1	1.1	1.1	1.1	1.1	1.1
Study Protocol Parameters Name
Study Protocol Parameters Name Term Accession Number
Study Protocol Parameters Name Term Source REF
Study Protocol Components Name	MTT; acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shaker	acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; biovision LDH-cytoxicity assay kit; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shaker	MTT; acetaminophen; dimethyl sulfoxide; glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shaker	M199 cell culture media; fetal bovine serum; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well plate adapter; orbital plate shaker	acetaminophen; M199 cell culture media; fetal bovine serum	triton-X-100; M199 cell culture media; fetal bovine serum	MTT;glycine; sodium chloride	acetaminophen;glycine; sodium chloride; triton-X-100; M199 cell culture media; fetal bovine serum; nanoparticle; costar 96 well flat bottom cell culture plates; plate reader; centrifuge set at 700-800 x g (Allegra X-15R, Beckman Coulter) with 96 well

plate adapter; orbital plate shakerbiovision LDH-cytoxicity assay kit

plate adapter; orbital plate shaker	biovision LDH-cytoxicity assay kit
Study Protocol Components Type	reagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrument

Study Protocol Components Type

reagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrument

reagent; reagent; reagent; reagent; reagent;reagent; reagent; reagent; material; instrument; instrument; instrument

reagent; reagent;material; instrument; instrument; instrument

reagent; reagent; reagent

reagent;

material; instrument; instrument; instrument

reagent; reagent

reagent;

reagent; reagent; reagent; reagent; reagent; reagent; material;

instrument; instrument; instrument

reagent

; reagent;material; instrument; instrument; instrumentreagent; reagent; reagentreagent; reagent; reagentreagent; reagent; reagentreagent; reagent; reagent; reagent; reagent; reagent; reagent; material; instrument; instrument; instrumentreagent


Study Protocol Components Type Term Accession Number	NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290;NPO_290; ; NPO_1436; NPO_1436; NPO_1436	NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290;NPO_290; ; NPO_1436; NPO_1436; NPO_1436	NPO_290; NPO_290;

Study Protocol Components Type Term Accession Number

NPO_290; NPO_290; NPO_290; NPO_290; NPO_290;NPO_290; ; NPO_1436; NPO_

290

1436; NPO_1436

NPO_290; NPO_290; ; NPO_1436; NPO_1436; NPO_1436

NPO_290; NPO_

1436

290; NPO_290

NPO_290; NPO_290; NPO_290

NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; NPO_290; ;

NPO_1436; NPO_1436; NPO_1436	NPO_290
Study Protocol Components Type Term Source REF	NPO; NPO; NPO

_290

; NPO

_290

;

NPO

_290

; NPO

_290

; NPO

_290

; NPO

_290

; NPO

_290

; ; NPO

_1436

; NPO

_1436

; NPO

_1436

NPO_290

NPO; NPO; NPO; NPO; NPO

_290

; NPO; NPO

_1436

;

NPO

_1436

; NPO

_1436NPO_290

; ; NPO; NPO

_290

; NPO

_290

NPO

_290

;

NPO

_290

;

NPO_290

NPO

_290

; NPO

_290

; NPO

_290

; NPO

_290

; NPO

_290

; NPO

_290

; ;

NPO

_290

; NPO

_290

; NPO

NPO

_290

; NPO

_290

; ; NPO

_1436

; NPO

_1436

; NPO

_1436

NPO_290Study Protocol Components Type Term Source REF

NPO; NPO; NPO

;

NPO; NPO; NPO; NPO

;

; NPO; NPO;

NPO

NPO;

NPO

; NPO; NPO; NPO

; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO; NPO; NPO; NPO; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO; NPO; ; NPO; NPO; NPONPO; NPO; NPONPO; NPO; NPONPO; NPO; NPONPO; NPO; NPO; NPO; NPO; NPO; NPO; ; NPO; NPO; NPONPO

NPO

Study Contacts

Example Investigation File Study Contacts Section

A	B
Study Person Last Name	Smith
Study Person First Name	Jane
Study Person Mid Initials	K
Study Person Email	smithj@mail.nih.gov
Study Person Phone	1231231235
Study Person Fax
Study Person Address	Laboratory Street, City, State 111111
Study Person Affiliation	Doe Laboratories
Study Person Roles	investigator
Study Person Roles Term Accession Number
Study Person Roles Term Source REF	MO

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Old Version 50

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Key

Ontology Source Reference

Ontology Source Reference

Investigation

Investigation

Publications

Investigation Publication

Investigation Contacts

Material

Study

Study

Study Design Descriptors

Study

Publications

Study Publications

Study Factors

Study Assays

Study Assays

Study

Protocols

Study Protocols

Study Contacts