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

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2024-03-04</periodOfReport>

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

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0000939568</rptOwnerCik>
            <rptOwnerName>KLOCK JOHN C JR</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>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>1</isOfficer>
            <isTenPercentOwner>0</isTenPercentOwner>
            <isOther>0</isOther>
            <officerTitle>Chief Executive Officer</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <nonDerivativeTable>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2024-03-04</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>2881140</value>
                </transactionShares>
                <transactionPricePerShare>
                    <footnoteId id="F1"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>2881140</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>I</value>
                    <footnoteId id="F2"/>
                </directOrIndirectOwnership>
                <natureOfOwnership>
                    <value>By John C. Klock, Jr. and Cynthia L. Klock Trust Dated 7/27/07</value>
                    <footnoteId id="F2"/>
                </natureOfOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
    </nonDerivativeTable>

    <footnotes>
        <footnote id="F1">Received in exchange for  8,407,114 shares of QT Imaging, Inc. (&quot;QT Imaging&quot;) in connection with the merger of QT Imaging into a subsidiary of QT Imaging Holdings, Inc. (the &quot;Company&quot;) pursuant to the exchange ratio set forth in the business combination agreement between QT Imaging and the Company. The closing price of the Company's Common Stock on the effective date of the merger was $3.53.</footnote>
        <footnote id="F2">The Common Stock are held directly by John C. Klock, Jr. and Cynthia L. Klock Trust Dated 7/27/07 (the &quot;Trust&quot;).  The Common Stock held by the Trust are beneficially owned jointly by Dr. John C. Klock, Jr., the Company's Chief Executive Officer and Director and Cynthia L. Klock, wife of Dr. John C. Klock, Jr., who jointly have sole voting and dispositive power over the shares held by the Trust.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ John C. Klock, Jr.</signatureName>
        <signatureDate>2024-03-06</signatureDate>
    </ownerSignature>
</ownershipDocument>
