<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0206</schemaVersion>

    <documentType>3</documentType>

    <periodOfReport>2024-03-21</periodOfReport>

    <noSecuritiesOwned>0</noSecuritiesOwned>

    <issuer>
        <issuerCik>0001844505</issuerCik>
        <issuerName>QT IMAGING HOLDINGS, INC.</issuerName>
        <issuerTradingSymbol>QTI</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001116229</rptOwnerCik>
            <rptOwnerName>GREENE JAMES S</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>C/O QT IMAGING HOLDINGS, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>3 HAMILTON LANDING, SUITE 160</rptOwnerStreet2>
            <rptOwnerCity>NOVATO</rptOwnerCity>
            <rptOwnerState>CA</rptOwnerState>
            <rptOwnerZipCode>94949</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>1</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>0</isTenPercentOwner>
            <isOther>0</isOther>
        </reportingOwnerRelationship>
    </reportingOwner>

    <nonDerivativeTable>
        <nonDerivativeHolding>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>200000</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>I</value>
                </directOrIndirectOwnership>
                <natureOfOwnership>
                    <value>By Sky D Ventures, LLC</value>
                    <footnoteId id="F1"/>
                </natureOfOwnership>
            </ownershipNature>
        </nonDerivativeHolding>
    </nonDerivativeTable>

    <footnotes>
        <footnote id="F1">The Common Stock is held directly by Sky D Ventures, LLC. The shares held by Sky D Ventures, LLC are beneficially owned by Mr. Greene, QT Imaging Holdings, Inc.'s member of the Board of Directors. Mr. Greene is also the Managing Member of Sky D Ventures, LLC, who has sole voting and dispositive power over the shares held by Sky D Ventures, LLC.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ James S. Greene</signatureName>
        <signatureDate>2024-04-03</signatureDate>
    </ownerSignature>
</ownershipDocument>
